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These samples (63 in total) have been aligned to the Ensembl 113 release of GRCh38 via subread, and counts have been generated using featureCounts with inbuilt hg38 genome

Data is located under 5FU Project on my computer All analysis saved in RDirectory Recovery_RNAseq

Individual 1 - 84-1 (9 samples)

Individual 2 - 87-1 (9 samples)

Individual 3 - 75-1 (9 samples)

Individual 4 - 75-1 (9 samples)

Individual 5 - 17-3 (9 samples)

Individual 6 - 90-1 (9 samples)

Individual 7 - 90-1 REP (9 samples)

Now that I have imported all of these counts files, I will make a matrix with all of the samples

First I’ll break them up by individual, and then combine them all together

Now that I’ve made individual matrices for each individual - I will combine them together into one large matrix

Version Author Date
a07d5bc emmapfort 2025-03-19

Version Author Date
a07d5bc emmapfort 2025-03-19
#Now that I've put together my matrix - I will do some QC checks

#now that I have all of my factors and my gene info, I will make some graphs to show the QC data

reads_by_sample <- c(tx_col)
fC_AllCounts %>% 
  ggplot(., aes (x = Conditions, y = Total_Align, fill = Treatment, group_by = Line))+
  geom_col()+
 geom_hline(aes(yintercept=20000000))+
 scale_fill_manual(values=reads_by_sample)+
  ggtitle(expression("Total number of reads by sample"))+
  xlab("")+
  ylab(expression("RNA-sequencing reads"))+
  theme_bw()+
  theme(plot.title = element_text(size = rel(2), hjust = 0.5),
        axis.title = element_text(size = 15, color = "black"),
        axis.ticks = element_line(linewidth = 1.5),
        axis.line = element_line(linewidth = 1.5),
        axis.text.y = element_text(size =10, color = "black", angle = 0, hjust = 0.8, vjust = 0.5),
        axis.text.x = element_text(size =10, color = "black", angle = 90, hjust = 1, vjust = 0.2),
        #strip.text.x = element_text(size = 15, color = "black", face = "bold"),
        strip.text.y = element_text(color = "white"))

Version Author Date
a07d5bc emmapfort 2025-03-19


####Read Counts by Treatment####
fC_AllCounts %>% 
  ggplot(., aes (x =Treatment, y= Total_Align, fill = Treatment))+
  geom_boxplot()+
 scale_fill_manual(values=reads_by_sample)+
  ggtitle(expression("Total number of reads by treatment"))+
  xlab("")+
  ylab(expression("RNA-sequencing reads"))+
  theme_bw()+
  
  theme(plot.title = element_text(size = rel(2), hjust = 0.5),
        axis.title = element_text(size = 15, color = "black"),
        axis.ticks = element_line(linewidth = 1.5),
        axis.line = element_line(linewidth = 1.5),
        axis.text.y = element_text(size =10, color = "black", angle = 0, hjust = 0.8, vjust = 0.5),
        axis.text.x = element_text(size =10, color = "black", angle = 90, hjust = 1, vjust = 0.2),
        #strip.text.x = element_text(size = 15, color = "black", face = "bold"),
        strip.text.y = element_text(color = "white"))

Version Author Date
a07d5bc emmapfort 2025-03-19

####Total Reads Per Individual####
fC_AllCounts %>% 
  ggplot(., aes (x =as.factor(Line), y=Total_Align))+
  geom_boxplot(aes(fill=as.factor(Line)))+
 scale_fill_brewer(palette = "Dark2", name = "Individual")+
  ggtitle(expression("Total number of reads by individual"))+
  xlab("")+
  ylab(expression("RNA-sequencing reads"))+
  theme_bw()+

  theme(plot.title = element_text(size = rel(2), hjust = 0.5),
        axis.title = element_text(size = 15, color = "black"),
        axis.ticks = element_line(linewidth = 1.5),
        axis.line = element_line(linewidth = 1.5),
        axis.text.y = element_text(size =10, color = "black", angle = 0, hjust = 0.8, vjust = 0.5),
        axis.text.x = element_text(size =10, color = "black", angle = 0, hjust = 1),
        #strip.text.x = element_text(size = 15, color = "black", face = "bold"),
        strip.text.y = element_text(color = "white"))

Version Author Date
a07d5bc emmapfort 2025-03-19

####Total Mapped Reads Per Drug####

reads_by_sample <- c(tx_col)
fC_AllCounts %>% 
  ggplot(., aes (x = Conditions, y = Assigned_Align, fill = Treatment, group_by = Line))+
  geom_col()+
 geom_hline(aes(yintercept=20000000))+
 scale_fill_manual(values=reads_by_sample)+
  ggtitle(expression("Total number of mapped reads by sample"))+
  xlab("")+
  ylab(expression("RNA-sequencing reads"))+
  theme_bw()+
  theme(plot.title = element_text(size = rel(2), hjust = 0.5),
        axis.title = element_text(size = 15, color = "black"),
        axis.ticks = element_line(linewidth = 1.5),
        axis.line = element_line(linewidth = 1.5),
        axis.text.y = element_text(size =10, color = "black", angle = 0, hjust = 0.8, vjust = 0.5),
        axis.text.x = element_text(size =10, color = "black", angle = 90, hjust = 1, vjust = 0.2),
        #strip.text.x = element_text(size = 15, color = "black", face = "bold"),
        strip.text.y = element_text(color = "white"))

Version Author Date
a07d5bc emmapfort 2025-03-19


####Read Counts by Treatment####
fC_AllCounts %>% 
  ggplot(., aes (x =Treatment, y= Assigned_Align, fill = Treatment))+
  geom_boxplot()+
 scale_fill_manual(values=reads_by_sample)+
  ggtitle(expression("Total number of mapped reads by treatment"))+
  xlab("")+
  ylab(expression("RNA-sequencing reads"))+
  theme_bw()+
  
  theme(plot.title = element_text(size = rel(2), hjust = 0.5),
        axis.title = element_text(size = 15, color = "black"),
        axis.ticks = element_line(linewidth = 1.5),
        axis.line = element_line(linewidth = 1.5),
        axis.text.y = element_text(size =10, color = "black", angle = 0, hjust = 0.8, vjust = 0.5),
        axis.text.x = element_text(size =10, color = "black", angle = 90, hjust = 1, vjust = 0.2),
        #strip.text.x = element_text(size = 15, color = "black", face = "bold"),
        strip.text.y = element_text(color = "white"))

Version Author Date
a07d5bc emmapfort 2025-03-19

####Total Reads Per Individual####
fC_AllCounts %>% 
  ggplot(., aes (x =as.factor(Line), y=Assigned_Align))+
  geom_boxplot(aes(fill=as.factor(Line)))+
 scale_fill_brewer(palette = "Dark2", name = "Individual")+
  ggtitle(expression("Total number of mapped reads by individual"))+
  xlab("")+
  ylab(expression("RNA-sequencing reads"))+
  theme_bw()+

  theme(plot.title = element_text(size = rel(2), hjust = 0.5),
        axis.title = element_text(size = 15, color = "black"),
        axis.ticks = element_line(linewidth = 1.5),
        axis.line = element_line(linewidth = 1.5),
        axis.text.y = element_text(size =10, color = "black", angle = 0, hjust = 0.8, vjust = 0.5),
        axis.text.x = element_text(size =10, color = "black", angle = 0, hjust = 1),
        #strip.text.x = element_text(size = 15, color = "black", face = "bold"),
        strip.text.y = element_text(color = "white"))

Version Author Date
a07d5bc emmapfort 2025-03-19

Now that I’ve made the final full matrix of all samples - 1. convert to log2cpm using cpm 2. filter for lowly expressed reads (rowMeans > 0)

fC_Matrix_Full
          DOX_24_Ind1 FLUO_24_Ind1 DMSO_24_Ind1 DOX_24rec_Ind1 FLUO_24rec_Ind1
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101928241           0            1            0              5               0
58155             526          709          717            910             525
1806               29           70           43             55              58
100873932           0            0            1              0               0
100873933           0            0            0              0               0
400765              6            5            7              7               2
100616452           3            1            0              2               1
406928              0            0            0              0               0
729987              7            1            5              9               2
51375             758          582          514            455             456
163404            372            0            0              3               0
100129620           1            0            0              4               0
9890               37           12            8             23              10
101928270           0            0            0              0               0
54873              11           12            4             25              15
391059            178          444          364            297             309
178              2283         4822         4604           2670            3185
23443             935         1226         1090           1172             840
64645            1580         2010         1687           1849            1513
163786            111          139          172            190              70
54482             176          406          412            617             347
127495            151          935          982            635             771
1629              995         1281         1075           1107             918
100506007          99           50           43            100              23
8634             1395         1772         1502           1848            1448
693138              0            2            0              4               1
8556              593          203          172            249             137
54112              59            3            3              7               2
100128787           0            0            0              0               0
7412              200         1911         1583            290            1231
2135             1326         1314         1173            666             978
148867           1421         1247          917           1344             967
51611             141          583          538            688             440
102606465          51          189          200            248             167
101928334           0            0            0              0               0
101928370         107          198          170            148             146
1901              797         7408         6446           3515            5628
104355286           0            0            0              0               0
118427              4            2            1              3               2
94236               1            0            0              1               1
1301              509          349          238            344             341
101928436           3            0            0              0               2
55599             511          730          661            780             617
280               789          528          453            611             445
648740             22           11           24             30              14
279                43           15           15             21               8
276                34           15           15             17               9
277                 3            0            1              1               3
278                 6           11            8              8               4
100129138           5            1            1              4               1
101928476           0            0            0              1               0
55170             582          718          727            839             573
22854              28           25           22             25              43
10451              58           13           14              8               6
102465839           0            0            0              0               0
100873946           0            0            0              0               0
29957            1439         1798         1577           1904            1442
148545              3            0            0              2               0
653149              0            0            0              0               0
284611            551          288          246            271             223
113802            102           39           50            120              35
55119            1560         1876         1716           2494            1471
163479             58            8            1             19               3
6814             1287         1408         1218           1874            1144
254268              5            0            0              5               0
642864              4            0            0              2               0
29899              32           45           50             41              18
23155            1649         2203         1826           1437            1604
22911            1404         1081          949           1041             873
6884              808          565          508            836             437
56900             920         1164         1212           1396            1050
677766             15            0            5              6               5
127003             23            0            2             10               0
57535             376           25           15            200              27
6301             2716         3311         2778           5069            2933
1952             2825         1550         1517           1319            1276
84722             105          105          102             72              53
343263            126          142          130            133             129
6272            20519        25394        20125          13337           19948
5686             1898         1652         1423           2459            1348
284612            876          661          615            583             457
127002            273          205          170            323             181
284613            243          398          356            621             335
57463             384          892          817            582             698
83873              39           15           22             18              18
2773              721         1384         1123           1901            1064
406974              3            4            5              0               0
2780               23           46           38             36              27
271              1302         1419         1199           1553            1233
2948              798          782          659            800             680
2946              562          454          380            307             404
2944               21           18           19             25              18
2949                7            7           12             11              10
2947             1635          775          623            781             688
79574               0            1            0              0               0
1435             2812          740          448           1242             470
10768            2913         2174         1953           2437            1695
85369            1258         1741         1519           1906            1435
257                13           22           18             33              19
104355139           5            0            0              0               1
164153              1            0            2              0               0
388662             98          203          149            222             160
100873974          50            1           10             34               2
3749             1059          948          821            588             773
440600             10            4            5              8               4
64783             769          381          317            511             243
9122               49           49           25             45              26
10542             826         1075          938           1555            1020
101410535          16            7            6             17               3
84432               2            7            1              4               5
643160              5            2            3              7               0
440602              7            2            0              7               1
3744                0            0            0              1               0
3737              214           14           16             20              15
3738              137            0            0              7               1
963                 3            1            1             12               1
55791             770          568          504            679             473
128338            455          896          844            848             676
10390             792          893          701            897             659
79961              25            2            6             24               2
1117               10            0            0              3               0
149620              4            0            0              0               0
27159               2            0            0              8               0
128344            180          173          211            225             150
441897              2            0            0              4               0
5016              269           38           21            154              25
79084            2023         1777         1486           2395            1402
515              4805         4953         4366           5166            4048
128346            157           21           30             60              19
57413              10            9            5              7               4
140                 1            1            0              1               0
100129269          95           54           37             33              53
5906             1427         2119         1707           2170            1816
55924            2141         2649         2367           1721            2238
100506343         112           68           61             48              49
101928718          80           51           28             13              26
11218             510          930          932            849             810
3752               71           91           60             44              57
100874295           0            0            0              0               0
100873995           0            0            0              0               0
643355             59            5           12             21               8
55917             858         1105          926           1454            1042
100422976           0            2            1              0               0
7482               72           41           22             57              24
54879             425          568          539            744             402
829              2674         3667         3301           5761            3261
4343             1632         1643         1395           1777            1448
389              5972         5750         4556           6992            5680
333926            602          102          107            200              68
284467             82            2            1              5               2
100996702          13           13            4              6               9
6566            10717        15096        13501           9020           11176
246182             12           21            6              7               4
100506392          15           34           29             46              24
100996251          13           10            8             22              12
9860              236          610          574            680             457
643441              0            0            0              0               0
260425            451          444          399            286             351
10745             510         1534         1237           1429             997
54665             496          402          381            446             320
100287722          32           19           10             31               9
26191              34            6            3              6               4
440603             16            8            6             17               7
10717             350          580          407            572             407
64858             497          114          106            129              87
101928846          41            5            1             12               7
204851           2583         1591         1471           2081            1155
56944             600          475          374            487             339
148281             22            0            0              2               1
51592             918         1619         1467           2032            1186
10286             572          803          749           1463             705
163259            150          111          134            172             102
270                12            0            0             10               0
4893             1201         1971         1856           2858            1552
7812            15474        14760        13001          16958           11825
80143            1284         1525         1335           1818            1192
6847                0            0            0              2               0
7252                3            4            0              0               0
10100             847         1062          975            635             652
4803              196          137           89            243             132
81839             895         1013          793            591             653
845             42041        32877        27861          13442           25878
4808              257            3            3            112               3
101928995           3            1            1              8               2
101928977           0            0            5              4               0
55356             368          888          777            709             771
126868             23           54           40             27              46
476             33928        44665        40324          31956           33935
84852              24           36           38             62              37
101929023           5            1            0              2               1
965               355          639          568            333             389
100616384           0            0            2              0               0
3321             6552         7013         6437           2349            5038
100302117           3            0            0              0               0
388667              0            0            0              0               0
914                 0            0            0              0               0
5738            14024        14994        13508           5618           11814
9398               47           32           30             26              29
101929099           5            0            3              3               1
8458              457          432          411            368             307
100126331           0            0            0              0               0
80263             364          161          199            252             153
79679              14            0            0              4               0
104355292           0            0            0              0               0
10905            1653         3621         3167           1925            2551
100996263           7            0            0              0               0
54855             516           59           52            187              46
54834             599          427          345            521             326
10885            1429         1137          961           1140             855
200162             14            1            0              8               2
6913                7            0            1              2               0
10352             248          270          276            262             197
104472716           2            0            0              0               0
101929147          81           90           86            111              94
51179               2            0            0              2               0
100874270           0            0            0              0               0
3284                1           17           17              0              12
3283                7            1            0              0               0
128102             18           26           17             24               6
644242             54           54           56             72              57
90874             792         1153         1093           1025            1020
26227            1283         1066          944            505             887
3158                0            0            0              2               0
83998               1            0            0              1               0
343505              4            0            0              6               0
11085               1            0            0              1               0
4853             8069        10418         8713           7626            7948
9554             1380         1819         1750           2161            1750
105378954          63           21           17             24              10
728841            709          740          684            909             657
767846             45           47           21             29              24
100996724        1039         2601         2372           1537            2152
101929796         194          187          154            178             147
101060684        1336         1071          971           1078             857
100189005           3            0            0              5               0
101954277           2            0            0              1               0
653505             39           15           14             15              13
728855            164           92           65            106              94
101954264           3            3            1              2               0
100189257           0            0            0              0               2
105369199           2            1            0              2               3
100996721           9            7           17              7               4
100126488           0            0            0              0               0
101929798           0            1            0              0               0
2210               15            3            4              7               2
100996318           7            1            3              2               0
337875              3            1            0              0               0
653820            319           92           82             80              44
653464            700         1146         1028           1144             897
100873165           5           10            9             17               8
647121             22           18           21             18              25
101929815           1            2            0              0               2
100189261           0            0            0              0               0
101954269           0            0            0              0               0
26863               0            1            0              0               0
791227              0            0            0              0               0
105369140           3            3            2              3               1
388692             11            6            4              4               2
100189142           0            0            0              0               0
100189326           0            0            0              0               0
199882              5            0            0              0               0
100189356           0            0            0              0               0
100132417          11            5            2              3               0
100996713           7            2            2              0               1
554282            250           50           68             56              35
100996712         444          608          551            632             522
101060524           2            0            0              0               0
730262              4            8            1              4               9
101954267           1            0            0              1               0
100189323           0            0            0              0               0
284565            718          785          673            803             685
100189127           2            0            0              2               0
728945              2            0            1              3               5
105371215           3            0            0              0               3
1818                3            0            0              0               0
645166              4           17            5             29              12
647135            641          957          798            961             723
728833            284           56           95             53              38
101929805           1            0            0              0               0
102724235          18           11           15              3               8
103091866          83           41           20             42              43
645142              1            2            1              0               0
101954266           2            2            1              8               2
7214                1            1            0              2               0
100288142         332          302          264            370             236
100126510           0            0            0              0               0
101060254          18           41           56             16              37
101929780         197          146          141            215             111
653519            384          465          461            363             334
5174               44           20           14             29              17
11126              22           45           45             50              28
27246             514         1502         1355           1597            1187
10623             597          696          563            892             572
200035            204           90          101            206             117
10401             873         1011          812           1016             904
102466191           0            0            0              0               0
148741             11            2            2              6               2
8515              331          396          326            121             291
8799              237          577          516            576             465
114814             12           25           26             18              27
9939             1583         1596         1368           2829            1289
128077            381          892          823           1065             732
84265             827          684          527            883             568
284615            132          394          337            346             316
10628            2034         4627         4069           4214            4355
148738             13           65           51             26              47
100189384           0            0            0              0               0
100189064           0            0            0              0               0
100188991           0            0            0              0               0
100189198           0            0            0              0               0
101954276           2            1            0              0               0
101928979          10           18           28             44              17
100132406         349          274          256            267             225
388677            229          260          281            352             226
170688            469         1438         1333           1231            1115
105371242           1            0            0              0               0
102724558           0            1            0              3               0
100288805           7            5            8             28               5
105373468          12           17           22              8              20
149013           1033          810          629            916             671
728989             20           59           34             27              32
100189334           0            0            0              0               0
101447996           0            0            0              0               0
644861              1            0            1              2               1
5565             3478         2084         1937           3043            1572
171423          14640        14165        12329           9112           10725
2330              107          132          103            137             101
9557              447          637          594            408             514
100289211           6           37           32             11              26
607               712          794          777            896             710
51205             255          455          412            503             404
2702              127         2002         2129            166            1864
2703                3            0            0              5               1
51463             447          572          550            440             412
100034743          40           17           30             46              11
100189140           2            0            0              0               0
26864               1            0            1              3               0
100189193           1            0            0              0               0
200030            401          319          285            383             276
101927468           0            0            0              0               1
100189056           0            0            1              0               0
100189307           0            0            0              0               0
100189174           0            0            0              0               0
100188998           0            0            2              0               0
100847044           0            0            0              1               0
101954273           0            0            0              1               0
102466225           0            1            0              0               0
26860               0            0            0              0               0
100132057          10           35           30             53              41
101954272           0            1            0              1               1
388685            197           72           72             79              50
644591              8            6            6              0               2
25832             953          842          699            878             551
100996763         399          381          335            397             235
440672            565         1855         1669           1583            1378
9659             3569         8862         8360           4665            6819
400818            465          461          376            522             368
653513            395          910          896            493             823
100996761          19           29           30             23              16
101060226         755          629          575            621             521
653598             41           18           24             34              37
57234             172           72           79            116              68
644634             15           14           12             12              13
100189228           0            0            0              0               0
102723955           5            1            2              0               5
100189059           1            0            0              0               0
440689            133           26           34             65              42
2209               54            7           19             27              16
101927767          24            0            8              9               3
653604             10            1            2              0               2
8370              109           36           47             74              59
105371405         351           88           86            178              97
126961             56            8            1             24               9
8337              451          108           67            443              91
337873             71           14           20             36              10
723790           1381          217          185           1594             247
333932             31            3            4              7               5
554313             52           13           20             18              15
8349             4306         2491         2263           4079            2093
8338               73            1            5             28               6
          DMSO_24rec_Ind1 DOX_144rec_Ind1 FLUO_144rec_Ind1 DMSO_144rec_Ind1
100287102               0               0                0                0
653635                198             166              197              251
102466751               0               5                3               13
100302278               2               0                0                0
645520                  0               0                0                1
79501                   0               0                0                0
729737                177             200              200              251
102725121               0               0                0                0
102723897             201             229              180              257
102465909               2               0                1                0
729759                  0               0                0                0
100132287             134             145              146              200
105378947               4               8                1                2
101928626               1               0                0                0
102465432            2456            2647             2606             3190
81399                   0               0                0                0
100133331             112             125              135              168
100288069              20               4               16               11
400728                  1               0                4                7
79854                  31              32               29               49
643837               1289            1169             1365             2134
284593                  0               0                0                2
100130417              22              30               23               31
148398                 13              30               28               32
26155                1603            1359             1371             1760
339451                445             412              367              576
84069                  16              17               14                8
84808                   8              11                3                8
57801                1029             641              716             1043
9636                  300             223              196              263
375790              23008           14589            16887            23764
105369174               8               7               10                2
105378948              18               2               16                8
401934                  1               0                0                2
54991                 248             231              278              316
254099                  0               0                0                0
406984                  0               0                0                0
406983                  0               0                0                0
554210                  0               0                0                0
254173                  0               0                0                0
8784                    1               2                0                1
7293                    0               0                2                4
51150                6872            6385             6318             8960
126792               1258            1153             1147             1738
388581                 19              23               21               41
118424                953             856              815             1192
6339                  187             186              217              261
116983               2029            1812             2029             2694
102465434               1               0                0                0
126789                204             184              200              269
54973                1209            1095             1044             1533
102465435               0               1                0                1
80772                 532             393              473              724
83756                  28              21               27               42
1855                 1163             992             1041             1524
102466740               0               0                0                0
54587                 640             568              552              763
54998                1083            1002              877             1432
81669                4970            4182             4005             5649
148413                307             259              277              374
55052                 971             890              736             1186
441869                  1               0                0                1
643965                  1               0                0                1
102724312               4               1                5                4
64856                 119              74               63               81
219293                 20              16               12               19
83858                 677             600              654              959
55210                 748             596              613              924
339453                 44              46               44               66
29101                1629            1746             1366             2060
643988                357             357              303              466
142678               1869            1725             1810             2393
8510                  524             473              445              772
984                   887             795              638             1077
728661               2338            2226             2200             3198
8511                  964            1077              941             1461
728642                785             742              689             1013
9906                 1012             975              932             1405
65220                1080             903             1065             1377
2782                 7034            5516             5854             9253
105378949               9              16               14               15
163688                  6              17               11               11
339456                  6               4                1                6
85452                   1               1                5                3
2563                    0               0                1                0
5590                  641             475              499              745
100506504              50              44               38               61
199990                692             624              593              850
105378592              10               9                6               24
6497                 1554            1065             1352             2132
105378594               0               0                2                6
79906                 105             109               84              128
100129534              21              23               23               26
11079                2675            2546             2514             3451
5192                  264             282              243              318
9651                   15              15               28               46
55229                 822             706              823             1080
388585                  0               2                0                2
115110                  0               0                0                0
8764                    0               0                0                0
100996583               2               2                0                2
127281                358             297              268              438
79258                   3               0                3                0
100287898              29              28               26               49
140625                  0               0                0                0
440556                 76              40               76               92
63976                 209             135              216              314
100422968               0               0                0                0
27237                 150              98              124              135
1953                 2924            2083             2373             3626
693135                  0               0                0                1
127262               1395            1202             1133             1886
49856                 252             229              217              320
7161                    6               8                8               16
57212                 777             777              801             1121
148870                 24              23               22               44
388588                 43              55               40               66
57470                1537            1263             1262             1915
9731                  674             579              642              908
1677                   80              82               60              122
339448                281             261              263              408
100133612              14              22               13               20
728716                  0               0                0                0
284661                  1               0                0                0
55966                   0               1                0                2
100616489               0               0                0                0
100616421               0               0                0                1
261734                205             127              180              203
8514                   52              61               53               65
26038                  20              39               52               72
6146                 2431            2360             1906             3166
102724450              15              15               21               34
388591               5389            4824             5771             7650
23463                2623            2688             2659             4087
148645                  2               0                4                3
390992                  0               0                0                0
387509                307             283              297              415
11332                 916             580              691             1036
54626                   2              14               12                9
83715                  11               4               15                4
100422975               0               0                0                0
8718                  577             450              542              618
57449                2530            2029             2231             2864
79707                 607             579              495              754
80835                   1               7                0                1
3104                  356             327              311              422
9903                 4268            3094             3525             5336
148479                588             512              553              848
90326                 187             210              197              255
55735                1036             871              976             1398
100505887               0               0                0                0
23261                 582             627              468              852
9341                 1824            1768             1277             2300
8863                  139             191              177              293
10911                   7               0                1                2
3604                   84              23               19               33
11315                2819            2638             2138             3694
54206                1769            1060             1218             1671
102724539               4               2                0                2
50651                  46              75               70               99
473                  3092            2825             3159             4368
102724552               3               0                1                4
2023                12230            9928             9856            13368
102465436               1               0                0                0
100505975               4              14                3               13
765                     0               1                0                0
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8338                   12               3                3                3
          DOX_24_Ind2 FLUO_24_Ind2 DMSO_24_Ind2 DOX_24rec_Ind2 FLUO_24rec_Ind2
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10905            2731         2613         2466           3355            2864
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54834            1076          369          322            754             435
10885             730          801          749            999             906
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6913                1            0            2              2               2
10352             381          419          389            405             422
104472716           0            1            0              0               0
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90874             510          505          437            680             552
26227            1135         1160         1201            234            1416
3158                2            0            0              5               5
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4853             9303         5522         5075           8022            6827
9554              728         1646         1549           2100            1922
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728841            331          485          426            463             478
767846              9           10            7              8               8
100996724        1495         4159         3066           1709            4206
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2210               32            8            4             30               1
100996318           6            1            6              4               3
337875              8            3            1              6               0
653820            230           56           50             39              99
653464           1013          667          613           1213             783
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647121              3            9           12             15              11
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100132417          47            3            4             65               3
100996713          12            7            4              8               4
554282            168           33           51             22              66
100996712         564          297          241            505             302
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647135            684          519          420            904             565
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653519            342          335          248            399             377
5174               18           18           12             17              16
11126               5           45           36             24              38
27246            1369         1976         1930           2397            2237
10623            1178          756          614            850             777
200035             84          149          138            146             132
10401             718          788          674           1049             879
102466191           0            0            0              0               0
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8515               24           55           55             11              84
8799              349          539          502            765             653
114814             11           37           28              7              34
9939             2967         2347         1960           4543            2374
128077            318          953          946           1211            1081
84265             959          794          673            587             766
284615            144          220          172            110             274
10628             226          406          504             82             796
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100132406         231          296          242            269             355
388677            325          479          459            577             526
170688            651         4934         3828           1987            5209
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105373468           7            3            5              6               8
149013            447          635          489            677             631
728989             16           37           33             15              42
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5565             5174         2851         2506           4734            3596
171423           4787         4604         4248           6060            4568
2330              217          134          131            194             133
9557              552          553          447            325             578
100289211           7           11           15              6               8
607               480         1159          971            898            1268
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2702               15         1269         1202             34            1599
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25832             657          907          744            672             976
100996763         381          623          533            727             684
440672            985         5872         5053           2467            6896
9659             6349        18149        14742           7860           18678
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653513            913         2238         1810           1068            2439
100996761          70           31           13             73              36
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57234             133           71           72            143              82
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440689            209           68           54            264              60
2209              242           37           31            235              36
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653604              5            1            2             12               2
8370              194           67           45            507              63
105371405         568          135           98           1649             134
126961            120           20           10            327              19
8337              758           78           62           1243              63
337873            106           19           15            282              19
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333932             51            2            3             64               9
554313             64           18           15            125              13
8349            11298         3877         3571          10972            3644
8338               50            5            6             45               6
          DMSO_24rec_Ind2 DOX_144rec_Ind2 FLUO_144rec_Ind2 DMSO_144rec_Ind2
100287102               5               6                3                9
653635                192             191              253              264
102466751               4               2                3                6
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26155                1808            1795             1913             2141
339451                183             225              250              266
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375790               5734           10430             5799             7364
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126792               1275            1147             1168             1379
388581                 22              17               13               31
118424                953             735              952              886
6339                  113             119              173              223
116983               1277            1328             1194             1270
102465434               0               0                0                0
126789                159             122              179              154
54973                1388            1206             1689             1722
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80772                 684             545              617              743
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1855                 1195            1096             1357             1477
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54998                1486             767             1228             1278
81669                2211            2980             3093             3412
148413                244             236              280              254
55052                1104             668             1079             1031
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64856                 148              74              100              123
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83858                 470             531              550              619
55210                 850             703              914             1002
339453                 71              38               81               84
29101                1989            1113             1706             1635
643988                721             368              497              515
142678               2238            2409             2735             2848
8510                  656             328              770              850
984                  1214            1004             1286             1397
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8511                 1286             619             1301             1369
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9906                  830             738             1026             1107
65220                 877             776              987             1047
2782                 7577            5454             5842             6139
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6497                 1476            1423             1489             1577
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79906                  96              92              163              204
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11079                2847            2550             2757             2931
5192                  328             268              254              276
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55229                1303            1194             1330             1408
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140625                  0               0                0                0
440556                580             251              920              925
63976                1381             789             1921             1734
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1953                  327             500              317              388
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7161                   16              22               10               13
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57470                2496            1896             2674             2852
9731                  839             888              899              987
1677                   68              52               79               89
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8514                   56              35               56               56
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388591               4517            3744             5772             6017
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3104                  369             386              429              464
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23261                1090             479              841              790
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8863                  369             187              441              526
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473                  3744            3508             4063             4351
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2023                30943           12154            20185            26767
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100506022              14               2                7               16
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199953               1177            1136             1208             1365
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22883                4658            6372             5722             6591
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10277                2632            1983             2729             2821
23095                7490            8987             8173             8762
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5226                 3119            2539             2534             2572
100526739               1               0                0                0
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1325                   16              11               15                8
1676                 1100             888             1104             1033
5195                  677             552              725              751
54897                5155            5305             5774             6507
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23435                3913            3236             3765             3746
10747                  53              76               60               85
6723                  724             382              562              546
5394                 1731            1439             1980             2011
2475                 4671            3979             4881             4831
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29914                 841             758              823              866
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93611                 808             781              945             1059
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284498                  1               0                1                0
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4524                 2581            1856             2857             3223
1185                 1282            1735             1781             2102
100379251              16              23                9                6
4878                68924           98668            40299            43035
4879                14760           35028             5319             6469
26829                   0               7                3                4
90231                2064            2017             1955             2214
5351                 7121            8940             4888             6314
9927                16177           21500            15466            16953
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6301                 3397            3006             5447             5365
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257                    27              38               24               29
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8349                 3572            4359             3749             3645
8338                    6               3                4                4
          DOX_24_Ind3 FLUO_24_Ind3 DMSO_24_Ind3 DOX_24rec_Ind3 FLUO_24rec_Ind3
100287102           2            0            0              0               0
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9636              185          166          206            348             163
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79739             479          352          336            291             324
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391051              7            0            1              6               0
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80135             529         1246         1020           1436            1004
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1486              503          419          428            521             420
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105378828          22           31           32            165              28
51100            1909         6376         5930           4232            5602
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2635                6           33           54             33              49
2633               14           11           10              5              15
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84146             841         1897         1853           4186            1908
164045             55           82           85             21              82
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7049              547          313          362            192             334
676                 3            0            5              6               2
253152             23           84           80             35              93
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7813              437          979          934            664             862
6125            12424        11444        10001           5151            8396
388650            231          569          573            157             642
6083                1            1            2              1               3
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22823             360          840          804           1050             713
50999            5153         5020         4436           2664            5250
343099            130          355          369            226             185
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1810             1233         1377         1315           2701            1286
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30836             724         1414         1228           2261            1332
2730              168          399          373            783             391
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5825             1437         3199         2990           1084            3118
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1266             3932         3825         3395           4307            3185
105378862          32           37           33             94              20
729970            163          179          161            200             147
199857             43          141          156             54             137
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148534           6953         6750         5941           1902            7942
100527978          41           82           72             21              95
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58155             442          930          852           1224             999
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406928              0            0            0              0               0
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51375             716          580          565            221             492
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100129620           6            0            0              1               0
9890               17           10           19             31              24
101928270           0            0            0              0               0
54873               3            9            8              6               2
391059            222          323          367            138             347
178              8231        10855         9969           1274           12877
23443             552          681          635            623             643
64645             701         1336         1382           1262            1518
163786             67          287          305             87             198
54482             166          371          404            238             367
127495            317         1013          920            271            1130
1629             1533         1987         1884           1325            1868
100506007          67           51           43             82              31
8634             1599         1464         1310            676            1319
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8556              573          236          199            379             210
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7412              527         5424         5150            102            5083
2135             1517         1131         1127            498            1157
148867            957          393          505            757             414
51611             120          787          674            181             680
102606465          84          212          202             61             208
101928334           0            0            0              0               0
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1901              617         6514         6207           1470            6236
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118427              4            0            8              9               2
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1301              344           92          117            322              99
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55599             685          813          889            582             764
280               867          611          715            402             462
648740             18           24           30             10              25
279                42           20           22             16              16
276                21           17           24             13               6
277                 6            6            1              1               1
278                18           19            9              5               5
100129138           0            4            0              1               3
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55170             535          954          955            745             786
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100873946           2            0            0              0               0
29957            1279         1717         1635           1112            1380
148545              1            0            0              0               0
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284611            523          645          604            339             379
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55119             559         1321         1309           1812            1328
163479             25            2            2             20               0
6814              695         1117         1056           1369            1193
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29899              43          192          166             47              81
23155            1112         1514         1524            614            1453
22911             854         1008          918           2948             877
6884              927          485          441           1077             439
56900             896         1645         1488            869            1735
677766             23            1            3             22               3
127003              7            1            3             25               1
57535             104            5            1            188              11
6301             1234         2595         2487           2875            2328
1952             2224         1532         1966           1213            1341
84722              46          179          210             38              85
343263             43           64           72             46              48
6272            11929        11917        12591           3074           10547
5686              800         1592         1441           1650            1308
284612            939          852          906            734             918
127002             95          132          166            353             123
284613             57          247          323            514             297
57463             187          779          826            262             957
83873              40           30           54             24              34
2773              254         1172         1080           1647            1105
406974              2            2            2              1               2
2780               16           12           25             20              29
271               771          932         1016            902             817
2948             1556         1174         1163            526             946
2946              911          585          605            272             395
2944               46           41           29             12              18
2949               22           12           18              5               8
2947             2379          938          794           2856             832
79574               0            0            0              1               0
1435              343          603          691           2326             628
10768            3278         2142         2217           2468            1940
85369             803         1398         1501           1151            1287
257                 6           23           32             18              11
104355139           0            0            0              1               0
164153              2            2            1              3               1
388662             58          159          195            109             203
100873974          14            4            1             40               6
3749              473          769          844            607             956
440600             21            5           14             16               1
64783             400          392          435           1236             341
9122               24           14           28            126              20
10542             934         1388         1195            803            1156
101410535          18            8           12             23              13
84432               3            1            2              3               0
643160              4            2            3              4               3
440602              7            2            3              2               5
3744                1            0            0              0               0
3737              124            2            4            118               7
3738               94            2            1             65               1
963                 1            0            0             10               0
55791             280          619          553           1181             699
128338            461         1046         1000            338            1174
10390             318          489          542            876             523
79961              11            5            2             82               5
1117               15            0            1            152               0
149620              4            0            0              2               0
27159               2            0            1              1               0
128344            227          271          262            146             228
441897              3            0            0              7               0
5016               89           10           18            225              17
79084            3613         1855         1772           1822            1600
515              7877         7155         6746           3056            7264
128346            128           50           44             90              58
57413               0            0            0              2               0
140                 0            0            0              2               0
100129269          29           24           20             21              11
5906              833         1822         1682           1288            1595
55924             983         2177         2095           2103            1917
100506343          28           45           25             62              24
101928718          45            7            5             28              11
11218             129          895          878            789             808
3752               35           44           54             22              49
100874295           1            0            0              0               0
100873995           0            0            0              0               0
643355             19            2            2              8              12
55917             164          872          833            952             821
100422976           0            0            0              0               0
7482               44           21           31             70              23
54879             249          501          478            408             522
829              3497         4087         3724           3451            3392
4343             1173          927         1229            900             781
389              4586         4524         4266           3628            3499
333926            326          103          117            247             105
284467             90            0            3             21               1
100996702           9            2            7              5               3
6566            14396        20124        20153          15117           23985
246182             14           18           20             41               8
100506392          22           25           43             27              27
100996251          14           17           19             14              10
9860               84          586          619            432             652
643441              1            0            0              0               0
260425            516          497          555            321             502
10745             423         1324         1485            581            1333
54665             667          519          496            711             541
100287722          23            4           12             26               7
26191              29            2            1             22               1
440603              8            0            1             15               1
10717             253          423          517            446             445
64858             320          187          201            276             124
101928846           8            6           10             34               8
204851           2283         2372         2417           4453            2013
56944             360          273          302             70             256
148281             36            0            0             83               0
51592             818         2185         2249           2021            1974
10286             209          934          819            952             840
163259            117          124          120            485             122
270                10            0            0              3               0
4893             1446         2008         1948           3264            1712
7812            30863        20400        18917          14836           18210
80143            1492         2002         1813           1180            1778
6847                0            0            0              0               0
7252                3            1            2              2               1
10100            1894         1485         1382            535            1979
4803                3           14            7            106               4
81839             179          841          810            498             661
845             41469        20445        19538           9857           30346
4808               55            4            2            580               4
101928995           1            0            0              1               2
101928977           1            0            1              2               1
55356             247          490          604            193             516
126868              7           48           29              6              20
476             35349        55994        61931          46393           51474
84852              11           70           47             86              44
101929023           2            2            1              4               0
965               202          496          433            197             578
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3321            10480         8575         9314           3454            7339
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388667              0            0            0              0               0
914                 2            0            0              3               0
5738            13196         8740         9881           2977            8478
9398               59           14           19             18              24
101929099           6            5            1              0               3
8458              286          450          466            476             319
100126331           0            0            0              0               0
80263             206          179          198            260             214
79679              18            0            0             11               0
104355292           0            0            0              0               0
10905            1623         3026         3029           1186            2939
100996263           1            0            0              0               0
54855             334           41           54           1417              36
54834             359          389          375            708             358
10885             839          939          896            900             941
200162              3            3            2              9               2
6913               33            2            3             42               0
10352             319          468          370            233             384
104472716           0            0            0              0               0
101929147          85          132          134            239             108
51179               0            2            0              1               0
100874270           0            0            0              0               0
3284                8            2            0              0               1
3283                6            0            0              2               0
128102              1            3            4             15               2
644242              4           14           10             53               7
90874              89          767          765            806             716
26227            1224          966         1112            523             922
3158                1            0            0             60               0
83998               0            0            1              5               0
343505              5            0            0              2               0
11085               2            0            0              1               0
4853             8080         8605         9614           7012            7391
9554             1521         2029         1914            927            1756
105378954          53           23           13             36               4
728841            760          643          765            502             538
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100996724        2587         4114         4649            882            3659
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653505             26           25            8             26               6
728855            168          105          112             62              83
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101929798           8            0            0              8               0
2210               17            7           12             82               8
100996318          14            2            1             14               6
337875             17            3            1             28               0
653820            295          109          107            111              72
653464            425          848          795            655             621
100873165           4            7           10              2               2
647121             17           10           23              4               9
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26863               3            0            1              3               1
791227              0            0            0              0               0
105369140           9            0            5              6               0
388692             17            4            7              3               6
100189142           0            0            0              0               1
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100132417          49            8            9             76               9
100996713          14            7            1             22               4
554282            308           85           71            106              38
100996712         339          400          356            511             286
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730262             14            6            6             11               1
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100189323           1            0            0              1               0
284565            627          562          792            415             594
100189127           1            0            2              1               0
728945              2            0            0              2               1
105371215          10            0            0              0               0
1818                2            0            0              0               0
645166              9            3            7              3               7
647135            467          620          645            673             409
728833            339           92           70            102              34
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102724235          18           21           18              7              13
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645142              3            1            0              1               0
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7214                0            0            2              7               2
100288142         313          236          338            230             221
100126510           0            0            0              0               0
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101929780         245          166          158            168             141
653519            212          332          345            275             375
5174               19           11           15             49              13
11126              22           40           44             44              35
27246             772         2576         2184           1631            2054
10623             406          625          647            777             651
200035            202          117           91            183             105
10401             554          843          887            793             881
102466191           0            0            0              0               0
148741             13            0            2             14               3
8515              361          340          310            101             244
8799              103          652          617            442             629
114814             16           37           25             28              20
9939             2392         2186         2170           2658            1692
128077            361          954         1065            390             851
84265             874          731          729            494             689
284615            111          191          212             69             197
10628             297         3195         3137           1262            3392
148738             19           45           42              2              49
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100132406         550          431          492            310             335
388677            552          584          588            289             469
170688            387         4087         3673           1068            3252
105371242           4            1            0              0               0
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100288805           9            5           18             10               9
105373468           6            8           24             11               6
149013            879          743          856            634             616
728989             42           83           91              9              48
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644861              0            0            0              3               0
5565             4997         3121         2587           4568            3226
171423           7730         6600         6579           4850            5758
2330              179          142          111            154             119
9557              869          554          664            266             482
100289211           4           36           22             32              19
607               531         1083         1219            848            1016
51205             308          600          607            241             490
2702               56         1223         1273             31             877
2703                7            0            0              3               0
51463             228          475          404            341             463
100034743          21           16            9             33              14
100189140           0            0            0              0               0
26864               0            0            0              1               0
100189193           0            0            0              0               0
200030            302          238          292            240             238
101927468           0            2            2              0               1
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100189307           0            0            0              0               0
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100188998           0            0            0              0               0
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102466225           0            0            0              0               0
26860               0            0            0              2               0
100132057          18           46           39             15              48
101954272           1            1            0              0               0
388685            172           53           52             80              46
644591             14            1            2              6               2
25832            1409         1129         1293            878             882
100996763         802          768          749            356             580
440672            462         5091         4563           1282            4166
9659            10182        18069        18125           4173           16115
400818            436          365          398            267             299
653513           1870         2472         2170            429            2069
100996761          20           30           37             32              48
101060226         937          809          929            712             603
653598             55           19           23             50              19
57234             189          117          114            129              77
644634             30           12           12              6               4
100189228           0            0            0              0               0
102723955           9            3            3              9               2
100189059           0            0            0              1               0
440689            173           87           67            279              43
2209              118           40           41            181              27
101927767          16           12           16             45               3
653604             16            3            0             18               1
8370               75           67           79            425              61
105371405         202          127          120           1153             124
126961             42            7            8            244              13
8337              279          115           99           1435              70
337873             34           17           24            198              17
723790            880          259          275           4595             205
333932             11            9            7            102               4
554313             27           14           17            130              24
8349            11996         4632         4183           6669            4493
8338               57            3            4             56               1
          DMSO_24rec_Ind3 DOX_144rec_Ind3 FLUO_144rec_Ind3 DMSO_144rec_Ind3
100287102               0               0                0                1
653635                162             243              272              176
102466751               1               5                6                7
100302278               0               0                0                0
645520                  0               2                0                0
79501                   0               0                0                0
729737                337             315              509              308
102725121               0               0                0                1
102723897             140             257              251              226
102465909               3               5                0                3
729759                  0               0                0                0
100132287             138             236              256              190
105378947               1               4                1                1
101928626               0               0                0                0
102465432            2132            3335             3345             1629
81399                   0               1                0                0
100133331             168             218              286              168
100288069              20              13               10               14
400728                  3               0                9                8
79854                  14              41               29               28
643837               2230             709             2604             2008
284593                  0               2                2                0
100130417              18              22               18               16
148398                 29              31               30               19
26155                1550            1586             1957             1488
339451                206             389              502              292
84069                   8              14                6                4
84808                   2               8                3                1
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          DOX_24_Ind4 FLUO_24_Ind4 DMSO_24_Ind4 DOX_24rec_Ind4 FLUO_24rec_Ind4
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163479             21            0            2              8               6
6814             1354         1301         1585           2041            2489
254268              3            0            0              3               0
642864              0            0            0              0               0
29899              33           64           82             35              50
23155             730         1472         1853           1086            2881
22911            2806          885         1085           1246            1541
6884              690          347          392            556             520
56900             601         1372         1504           1422            2280
677766              6            5            7              5              10
127003             15            4            0              4               0
57535             216           13           14             41              18
6301             2745         2378         2836           5299            4215
1952             5845         1383         2113           1191            2382
84722              28           44           86              8              60
343263             13           15           20             43              30
6272             7295         9650        13605           5322           20027
5686             1751         1458         1766           2539            2412
284612           1957         1124         1222           1153            1910
127002            295          204          175            238             279
284613            248          372          502            960             650
57463             273          620          748            526            1106
83873              30           57           45             21              75
2773              820         1039         1598           2782            2090
406974              1            1            3              0               0
2780               18           17           38             22              35
271              1316          972         1331           1315            1751
2948              697         1035         1144            754            1575
2946              323          482          481            259             829
2944               12           15           19              7              24
2949                6           18           11             12              12
2947             2222          815          773            601            1198
79574               0            0            0              1               1
1435             3491         1323         1371           3026            2093
10768            3465         2046         2479           2768            3771
85369            1577         1589         1844           2244            2792
257                11           17           42             48              36
104355139           3            0            1              1               0
164153              3            0            2              0               2
388662             97          320          297            193             433
100873974         168            2            2              6               0
3749             1920          632          575            448             796
440600             17            7            4             19               6
64783            1210          342          413            328             495
9122                8           11           30             21              48
10542             485         1332         1273           1620            2004
101410535          23           14           16             16              24
84432               2            1            5              1               8
643160              2           16            8              6               7
440602              6           22           10             14              13
3744                0            0            0              2               0
3737              651           24           24             15              39
3738              194            0            0              1               0
963                 2            0            0              2               0
55791            1023          602          647            707             902
128338            400         1076         1207            984            1914
10390             771          632          749           1089            1116
79961              88            5           11             36              17
1117               20            0            0             13               2
149620              1            0            0              0               0
27159               0            0            0              1               1
128344            178          238          187            266             312
441897              1            0            0              0               0
5016              255           21           27             22              24
79084            1648         1402         1669           2629            2251
515              3357         6173         5869           5836            9446
128346             83           59           46             45              73
57413               0            0            0              1               1
140                 1            0            0              0               0
100129269         104           16           25             21              19
5906             1157         1445         1614           2217            2462
55924            3732         1835         2179           1676            3074
100506343         135           45           37             45              54
101928718          86           16           19              7              13
11218             679          747          888            840            1329
3752               76           29           21              6              35
100874295           0            0            0              0               0
100873995           0            0            0              1               0
643355             20            2            1              2               2
55917             580          891         1262           1504            1628
100422976           0            0            0              0               2
7482               79           17           25             26              27
54879             345          393          431            616             647
829              2362         3512         4235           6995            6302
4343             1680         1170         1529           1948            2256
389              3633         4507         6088          10756            8783
333926            709           83           99            316             145
284467            154            0            0              2               0
100996702           6           10           12             15              13
6566             9908        23777        24297          10235           42639
246182             10           18           14             13              14
100506392          17           52           34             50              66
100996251          22           15            7             24              22
9860              158          642          589            774             996
643441              0            0            0              1               0
260425           1197          496          670            340             957
10745             727         1170         1591           1644            2165
54665             590          490          575            557             889
100287722          27           13            9              9              18
26191              31            2            5              4               3
440603              4            0            3              5              11
10717             429          377          395            480             597
64858             474          129          189             90             245
101928846          65            2            6              2              10
204851           4066         1792         2295           2509            3124
56944             326          352          373            203             623
148281             17            0            0              3               0
51592            1337         2066         2318           2618            3368
10286             342          741          783           1628            1199
163259            269          138          144            298             211
270                 6            0            0              1               0
4893              993         1730         2156           2942            2899
7812            17840        16515        18048          24160           26742
80143            1041         1687         1955           2303            2624
6847                1            0            0              7               0
7252                1            4            2              0               1
10100             420         1184         1160            530            1916
4803               74           28           33            193              99
81839             857          700         1016            522            1353
845              8539         7850         6945           2551           14038
4808              535            4            2             79               5
101928995           3            1            1              1               0
101928977           1            0            0              0               2
55356             241          565          574            329             903
126868             18           15           25             10              33
476             30468        55183        65536          44977          106428
84852              20           44           52            102              63
101929023           3            0            0              1               5
965               199          445          574            415            1042
100616384           0            0            0              0               0
3321             8376         6804         8768           3812           11904
100302117           0            0            0              0               0
388667              0            0            0              0               0
914                 2            0            0              0               0
5738            10450        13194        17519           4491           25593
9398               55           24           41             20              57
101929099           7            1            0              3               2
8458             1206          403          562            692             660
100126331           0            0            0              0               0
80263             297          278          281            322             412
79679              21            0            0              1               0
104355292           0            0            0              0               0
10905            2362         3127         4100           3059            6009
100996263           2            0            0              1               0
54855             625           25           31            330              25
54834            1243          401          528            732             666
10885            1683          851         1201           1450            1706
200162              3            3            5              9               2
6913               16            2            2              7               2
10352             281          310          371            378             614
104472716           0            0            0              1               0
101929147         112           95          101            102             143
51179               1            0            0              2               0
100874270           0            0            0              0               0
3284                2            7           11              6              16
3283               12            0            0              2               0
128102              6            1            3             23               3
644242             19            2           13             58               6
90874             779          754          817            769            1286
26227             723          581          894            289            1104
3158                0            0            0              0               0
83998               0            0            0              0               0
343505              1            1            0              2               0
11085               0            0            0              0               0
4853            11556         8025        11607           7561           13940
9554              669         1935         2305           2829            3379
105378954          88           23           22             47              26
728841            483          619          806            829            1131
767846             23           15           16             11              16
100996724        1379         4333         3897           1451            6215
101929796         155          201          221            165             321
101060684         771          832          967            801            1347
100189005           4            0            0              3               0
101954277           1            0            3              1               0
653505             11            9           13             39              22
728855            155           93           99            103             131
101954264           0            0            0              0               0
100189257           0            0            0              0               0
105369199           2            7            1              3               2
100996721           9            6           10              1               6
100126488           0            1            0              0               1
101929798           0            0            0              1               0
2210               25            7           13             42              11
100996318          20            4            3              7               5
337875              6            1            3              7               2
653820            448           42           71             29              36
653464            949          480          679           1097            1032
100873165           1            2            2              3               4
647121              5           16           27             18              39
101929815           0            1            2              2               4
100189261           0            0            0              0               0
101954269           0            0            0              1               0
26863               1            0            0              2               3
791227              0            0            0              0               0
105369140           1            2            2              0               5
388692              5            4            4              1               1
100189142           0            0            0              0               0
100189326           0            0            0              0               0
199882              0            0            2              0               0
100189356           0            0            0              0               0
100132417          39           10            6             35              12
100996713           6            4            6             12               7
554282            303           28           33             20              35
100996712         671          256          374            454             463
101060524           1            0            0              2               1
730262              3            1            4             11               1
101954267           0            2            0              0               1
100189323           0            1            0              0               0
284565            475          549          748            474             886
100189127           0            0            0              0               1
728945              1            0            0              1               1
105371215           2            0            0              0               0
1818                0            0            0              1               0
645166             17           32           25             35              58
647135            741          368          492            696             670
728833            369           23           53             21              22
101929805           1            0            1              0               0
102724235          13            8           15              7              12
103091866         104           19           26             26              47
645142              1            0            0              0               0
101954266           3            0            0              2               1
7214                0            0            0              2               0
100288142         214          205          247            229             312
100126510           0            0            0              0               0
101060254          16           40           34             19              52
101929780         215          110          175            183             200
653519            271          306          386            361             614
5174               23            9           10             21              20
11126              22           26           39             37              54
27246             614         1668         1959           1617            3042
10623             840          637          698           1025            1099
200035            214          118          115            234             166
10401             907          876         1038           1384            1584
102466191           0            1            0              0               0
148741              6            3            0              3               1
8515               97          156          172             54             310
8799              247          601          638            745            1001
114814              4           35           30             23              49
9939             1933         1657         1894           4391            2588
128077            233          736         1004           1214            1428
84265             852          538          568            639             915
284615            200          178          210            129             304
10628             766         4772         6892           5264           14227
148738             10           50           35              7              63
100189384           0            0            0              0               0
100189064           1            0            0              0               0
100188991           0            0            0              0               0
100189198           0            0            0              0               0
101954276           0            0            1              0               1
101928979           7           40           33             31              35
100132406         308          317          403            278             434
388677            272          362          431            315             543
170688            609         3984         4196           2772            6754
105371242           0            0            0              0               0
102724558           0            2            3              1               3
100288805           6           12           14             13               9
105373468           7           15           18              7              25
149013            681          657          839            838            1142
728989             21           49           50             19              81
100189334           0            0            0              0               0
101447996           0            0            0              0               0
644861              0            0            0              3               2
5565             3752         2327         2229           2971            3683
171423           7111         7944         9772           9298           15186
2330              158          122          146            224             222
9557              496          536          672            324             865
100289211           0           15           10              3              18
607               700         1078         1263           1199            1933
51205             193          407          406            237             629
2702                9          630          977             39            1642
2703                6            0            0              3               0
51463             300          400          454            388             793
100034743          13           15           14             16              23
100189140           0            0            0              0               0
26864               0            0            0              0               0
100189193           0            0            0              0               0
200030            267          281          349            323             432
101927468           0            0            0              1               0
100189056           0            0            0              0               0
100189307           0            0            0              0               0
100189174           0            0            0              0               0
100188998           0            0            0              0               1
100847044           0            0            5              0               5
101954273           0            1            1              0               6
102466225           0            0            1              0               0
26860               1            0            0              0               0
100132057           4           34           28             23              49
101954272           0            6            0              1               0
388685            194           47           80             96             115
644591              4            3            6              4               3
25832             619          721          814            501             961
100996763         366          436          507            373             638
440672            805         5043         5296           3418            8806
9659             5689        16873        16137           5066           24700
400818            278          368          445            350             607
653513            467         1599         1440            476            2464
100996761          37           26           31             28              47
101060226         549          572          692            488             847
653598             14           13           30             64              46
57234             183           96          117            143             168
644634             20            3           15              4              15
100189228           0            0            0              0               0
102723955           0            2            0              0               1
100189059           0            0            0              0               0
440689            200           75           51            259             102
2209              202           40           37            265              40
101927767          39           12            8             68              17
653604             11            0            2             16               7
8370              186           56           64            506             100
105371405         529          136          151           1683             200
126961            130           23           18            300              21
8337              708          100           87           1400             120
337873             96           23           23            283              31
723790           2121          277          251           3299             335
333932             55            5            4             69               4
554313             37           13           19            127              27
8349             8874         4357         3968          13125            6510
8338               63            1            3             37               7
          DMSO_24rec_Ind4 DOX_144rec_Ind4 FLUO_144rec_Ind4 DMSO_144rec_Ind4
100287102               1               6                0                2
653635                311             368              351              297
102466751              12              17               16                4
100302278               1               0                0                0
645520                  0               0                0                0
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729737                396             391              400              387
102725121               3               5                3                0
102723897             359             401              401              327
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729759                  0               0                0                0
100132287             174             188              175              177
105378947               4               2                1                0
101928626               0               0                0                0
102465432            1755            2171             1784             1890
81399                   0               0                0                1
100133331             140             196              153              173
100288069              34              18               30               36
400728                  2               3                2                3
79854                  38              32               28               34
643837               1899            1329             2413             2144
284593                  2               1                2                0
100130417              22              16               17               22
148398                 10               8               19               17
26155                1699            1624             1491             1441
339451                350             349              298              292
84069                  15               7                6               11
84808                   7               5                2                1
57801                1161             964              909             1008
9636                  144             123              111              137
375790              14759           16610            11017            14190
105369174               6               6                9               14
105378948              14               9               13               23
401934                  2               0                0                0
54991                 240             216              248              273
254099                  0               0                0                0
406984                  0               0                0                0
406983                  0               0                0                0
554210                  0               0                0                0
254173                  0               0                0                0
8784                    4               2                0                3
7293                    0               0                0                0
51150                6852            7767             5985             6163
126792               1535            1481             1306             1298
388581                 14              18               13               12
118424                940             854              816              769
6339                  133             116              146              132
116983               1663            1951             1621             1490
102465434               0               0                0                0
126789                213             196              189              185
54973                1364            1368             1334             1221
102465435               1               0                0                0
80772                 589             538              518              555
83756                  30              57               37               40
1855                 1216            1169             1225             1154
102466740               0               0                0                0
54587                 735             511              492              552
54998                1060             958             1004              879
81669                4085            4225             3772             3973
148413                261             236              258              217
55052                1033             842              948              885
441869                  0               0                1                0
643965                  2               2                4                0
102724312              17              12                8               21
64856                 670             234              375              656
219293                 51              73               40               42
83858                 748             812              647              621
55210                 744             751              655              708
339453                 67              47               63               56
29101                1608            1220             1305             1254
643988                146             128              156              190
142678               2108            2438             2088             2000
8510                  548             519              608              473
984                  1041            1020              987              983
728661               2405            2200             2197             2225
8511                 1059             970             1189              910
728642                811             834              720              692
9906                 1477            1362             1470             1391
65220                1120             988              889              925
2782                 8204            6062             6490             7735
105378949              16              11                5               10
163688                  4               7                4                3
339456                  2               2                6                2
85452                   1               2                6                6
2563                    1               0                2                2
5590                  772             613              749              580
100506504              35              50               37               43
199990                699             476              515              563
105378592               9              11                7                8
6497                 1690            1681             1555             1747
105378594               3               3                1                0
79906                  90              69               82               81
100129534              22              15               13               19
11079                3215            3318             2710             2801
5192                  328             263              258              233
9651                    8              15                6               17
55229                1155            1297             1244             1163
388585                  0               0                0                0
115110                  0               0                0                0
8764                    0               3                0                2
100996583               0               0                0                0
127281                585             357              559              529
79258                   1               2                3                2
100287898              17              14                6               12
140625                  0               0                0                0
440556                483             145              561              459
63976                1072             743             1295             1276
100422968               0               0                0                0
27237                 100              43               88               83
1953                 1606            1021             1082             1541
693135                  0               0                0                0
127262               1316            1078             1146             1277
49856                 289             262              254              243
7161                   10               7                4                7
57212                 971             831              907              909
148870                 34              28               32               30
388588                 27              26               28               31
57470                1898            1708             1903             1768
9731                 1028             962              941              998
1677                   71              78               63               79
339448                251             244              278              254
100133612               7               8                2                4
728716                  0               0                0                0
284661                  0               0                0                0
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100616421               0               0                0                0
261734                198             157              195              189
8514                   58              45               89               68
26038                  16              33               18                9
6146                 2462            1527             2179             2240
102724450              39              46               27               21
388591               5343            4866             5346             4798
23463                3712            3645             3270             3436
148645                  7               2                1                1
390992                  0               0                0                0
387509                261             225              230              243
11332                 637             444              458              608
54626                   2               8                0                5
83715                   7              14               15                9
100422975               0               0                0                0
8718                  243             320              190              178
57449                1022            1415              804              758
79707                 750             691              622              676
80835                   1               2                1                4
3104                  387             387              362              318
9903                 3475            3834             2812             2959
148479                937             690              900              832
90326                 189             159              173              178
55735                1465            1223             1398             1348
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23261                 783             515              709              629
9341                 2650            1750             2101             2368
8863                  264             157              484              471
10911                   0               1                2                5
3604                  293             109              107              247
11315                3308            2589             2892             2916
54206                1262            1153              746             1036
102724539               0               0                0                0
50651                  46              87               57               75
473                  3784            4003             3557             3783
102724552               0               1                0                1
2023                23793           13359            30130            35783
102465436               0               0                1                0
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765                     0               2                0                0
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6518                    1               3                2                8
80045                  81              62               64               62
407040                  4               4                3                1
9563                 2726            2525             2798             2741
80176                 146             150              110              154
100506022              19              13                6               14
84275                 154             106              127              142
199953                978             883              998              938
5293                  174             187              197              191
644997                  0               1                0                0
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22883                6784            7193             6210             6700
56998                 710             546              703              653
84328                 660             475              599              648
64802                 823             732              836              747
100847055               0               1                0                1
105376719               1               0                1                0
116362                  3               6                3                0
10277                2518            2059             2383             2399
23095                6952            7317             6578             6948
100422970               1               0                0                0
5226                 3270            2958             2737             2690
100526739               0               0                0                2
378708                 29              16               37               36
1325                    4               8                6                6
1676                  846             716              768              817
5195                  573             654              539              522
54897                4253            4639             4001             4010
148345                 47              32               32               22
23435                3801            3316             3393             3510
10747                  73              87               85               62
6723                  878             538              647              743
5394                 1758            1652             1690             1659
2475                 4143            3830             3929             4176
100873935               0               2                0                0
10218                   3              13               16               14
29914                 771             626              690              712
57540                   6               5                0                7
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26232                 214             230              141              135
93611                 869             829              880              766
26270                 114              86              122              113
10459                 395             225              342              300
374946                  9               7                8                1
57085                 428             456              359              420
284498                  0               0                0                0
102724659               1               0                0                0
4524                 2435            2175             2634             2494
1185                 1551            1864             1545             1589
100379251               7              21                6                8
4878                66249          100302            49985            53451
4879                13575           33896             8860            12978
26829                   2               1                0                1
90231                2302            2549             2264             2347
5351                10062           10591             9522            10518
9927                12089           14011            11552            12192
60672                 361             556              310              357
102466982               0               0                0                0
943                     0              15                2                2
102465836               0               0                0                0
7133                    4               8                2                8
100616438               0               0                0                0
55187                2675            3045             2834             3364
677885                  4               8                6                9
9249                  400             308              288              511
102466722               0               0                2                0
343066                  0               1                0                0
126767                  0               0                0                0
93190                   0               0                0                0
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27246             436         1765         1566           1418            1696
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8799              196          648          511            613             578
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9939             1774         1611         1525           2881            1548
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284615            156          248          217            157             214
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105373468          11           24           24             17              42
149013           1127         1075          907           1006            1380
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5565             4501         3251         2731           5186            2824
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2330              157          141          141            173             179
9557              626          869          764            457             834
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607               808         1088          822           1124            1096
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2209              109            7           22             79              10
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554313             56           17           11             40              12
8349             5498         2909         2635           6253            2672
8338               69            4            4             26               2
          DMSO_24rec_Ind5 DOX_144rec_Ind5 FLUO_144rec_Ind5 DMSO_144rec_Ind5
100287102               9               7                6                6
653635                177             182              158              180
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          DOX_24_Ind6 FLUO_24_Ind6 DMSO_24_Ind6 DOX_24rec_Ind6 FLUO_24rec_Ind6
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8349            11128         3510         3238           8816            3280
8338               82            6            2             35               7
          DMSO_24rec_Ind6 DOX_144rec_Ind6 FLUO_144rec_Ind6 DMSO_144rec_Ind6
100287102               5               4                4                9
653635                247             225              254              251
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          DOX_24_Ind6REP FLUO_24_Ind6REP DMSO_24_Ind6REP DOX_24rec_Ind6REP
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8338                        2                   3
 [ reached getOption("max.print") -- omitted 26808 rows ]
fC_Matrix_Full_cpm <- cpm(fC_Matrix_Full, log = TRUE)

boxplot(fC_Matrix_Full_cpm, xlab = "Conditions", ylab = expression("log"[2]* "counts per million"), ylim= c(-10, 20))

Version Author Date
a07d5bc emmapfort 2025-03-19
hist(fC_Matrix_Full_cpm, xlab = expression("log" [2]* "counts per million"))

Version Author Date
a07d5bc emmapfort 2025-03-19

#first check the number of variables in the cpm file
dim(fC_Matrix_Full_cpm)
[1] 28395    63
#[1] 28395    63

#now use rowMeans to filter out values with mean < 0 (this means rowMeans > 0 is what I'm left with) for each row/gene in the original file
rowMeans_All <- rowMeans(fC_Matrix_Full_cpm)
fC_Matrix_Full_cpm_filter <- fC_Matrix_Full_cpm[rowMeans_All > 0,]
dim(fC_Matrix_Full_cpm_filter)
[1] 14225    63
#[1] 14225    63


#with this I've filtered out a significant number of rows, want ~14000

hist(fC_Matrix_Full_cpm_filter, 
     main = "Recovery All Individuals log2cpm Filtered", 
        xlab = expression("log"[2]*"counts per million"))

Version Author Date
a07d5bc emmapfort 2025-03-19
boxplot(fC_Matrix_Full_cpm_filter, 
        main = "Recovery All Individuals log2cpm Filtered", 
        xlab = "Conditions", 
        ylab = expression("log"[2]*"counts per million"),
        ylim = c(-10,15))

Version Author Date
a07d5bc emmapfort 2025-03-19

#save this to an R object
#saveRDS(fC_Matrix_Full_cpm_filter, "data/fC_Matrix_Full_cpm_filter.RDS")
#this will be used to filter my datasets against for filtered counts
#through this, I can save the rownames to have my final filtered dataset for DE

filt_gene_list <- row.names(fC_Matrix_Full_cpm_filter)
#saveRDS(filt_gene_list, "data/filt_gene_list.RDS")

#this is what I'll be using for differential expression analysis (see DE page)
####histogram of total counts, unfiltered####
hist(fC_Matrix_Full_cpm, main = "Recovery All Individuals log2cpm Unfiltered",
     xlab = expression("log" [2]* "counts per million"))

Version Author Date
a07d5bc emmapfort 2025-03-19
####histogram of total counts, filtered####
hist(fC_Matrix_Full_cpm_filter, 
     main = "Recovery All Individuals log2cpm Filtered rowMeans > 0", 
        xlab = expression("log"[2]*"counts per million"))

Version Author Date
a07d5bc emmapfort 2025-03-19
####boxplot log2cpm all samples####

boxplot(fC_Matrix_Full_cpm, xlab = "Conditions", ylab = expression("log"[2]* "counts per million"), ylim= c(-10, 20), main = "Boxplots of log2cpm per sample")

Version Author Date
a07d5bc emmapfort 2025-03-19
####boxplot log2cpm filtered all samples####

boxplot(fC_Matrix_Full_cpm_filter, xlab = "Conditions", ylab = expression("log"[2]* "counts per million"), ylim= c(-10, 20), main = "Boxplots of log2cpm Filtered rowMeans > 0 per sample")

Version Author Date
a07d5bc emmapfort 2025-03-19

Now that I’ve filtered the samples after log2cpm conversion, I can start to make heatmaps and PCA plots to look at my data. I want these plots to have treatment and time as the primary variables.

PCA_data <- fC_Matrix_Full_cpm_filter %>% 
  prcomp(.) %>% 
  t()

PCA_data_test <- (prcomp(t(fC_Matrix_Full_cpm_filter), scale. = TRUE))


#ind_num <- c("1", "1", "1", "1", "1", "1", "1", "1", "1","2", "2", "2", "2", "2", "2", "2", "2", "2", "3", "3", "3", "3", "3", "3", "3", "3", "3","4", "4", "4", "4", "4", "4", "4", "4", "4","5", "5", "5", "5", "5", "5", "5", "5", "5","6", "6", "6", "6", "6", "6", "6", "6", "6","6R", "6R", "6R", "6R", "6R", "6R", "6R", "6R", "6R")

#saveRDS(ind_num, "data/ind_num.RDS")

ind_num <- readRDS("C:/Users/emmap/RDirectory/Recovery_RNAseq/Recovery_5FU/data/ind_num.RDS")

####now make an annotation for my PCA####
annot <- data.frame("samples" = colnames(fC_Matrix_Full_cpm_filter)) %>% separate_wider_delim(., cols = samples, names = c("Tx", "Time", "Ind"), delim = "_", cols_remove = FALSE) %>% unite(., col = "Tx_Time", Tx, Time, sep = "_", remove = FALSE) %>% cbind(., ind_num)

#saveRDS(annot, "C:/Users/emmap/RDirectory/Recovery_RNAseq/Recovery_5FU/data/annot.RDS")


#combine the prcomp matrix and annotation
annot_PCA_matrix <- PCA_data_test$x %>% cbind(., annot)


#now I can make a graph where I have filled values for individual! I have seven colors for seven individuals
# I have three fill values for three timepoints

#using annotation matrix above as well as annotated PCA matrix (annot_PCA_matrix)

#extra info for colors in the graph (fill parameter)
fill_col_ind <- c("#66C2A5", "#FC8D62", "#1F78B4", "#E78AC3", "#A6D854", "#FFD92A", "#8B3E9B")

fill_col_ind_dark <- c("#003F5C", "#45AE91",  "#58508D", "#BC4099", "#8B3E9B", "#FF6361", "#FF2362")

fill_col_tx <- c("#63666D", "#499FBD", "#DCACED")

#I want the color parameter to be treatment, the shape as time, and individual as number

####PC1/PC2####
annot_PCA_matrix %>% ggplot(., aes(x=PC1, y=PC2, size=10)) +
  geom_point(aes(color = Tx, shape = Time)) +
  scale_color_manual(values = c(fill_col_tx))+
  scale_shape_manual(values = c(15, 19, 17))+
  geom_text_repel(aes(label = ind_num))+
  theme_bw(base_size = 10)+
  ggtitle(expression("PCA of filtered log"[2]*"cpm"))+
  guides(size="none")
Warning: ggrepel: 10 unlabeled data points (too many overlaps). Consider
increasing max.overlaps

Version Author Date
a07d5bc emmapfort 2025-03-19
  

####PC2/PC3####
annot_PCA_matrix %>% ggplot(., aes(x=PC2, y=PC3, size=10)) +
  geom_point(aes(color = Tx, shape = Time)) +
  scale_color_manual(values = c(fill_col_tx))+
  scale_shape_manual(values = c(15, 19, 17))+
  geom_text_repel(aes(label = ind_num))+
  theme_bw(base_size = 10)+
  ggtitle(expression("PCA of filtered log"[2]*"cpm"))+
  guides(size="none")
Warning: ggrepel: 7 unlabeled data points (too many overlaps). Consider
increasing max.overlaps

Version Author Date
a07d5bc emmapfort 2025-03-19

####PC3/PC4####
annot_PCA_matrix %>% ggplot(., aes(x=PC3, y=PC4, size=10)) +
  geom_point(aes(color = Tx, shape = Time)) +
  scale_color_manual(values = c(fill_col_tx))+
  scale_shape_manual(values = c(15, 19, 17))+
  geom_text_repel(aes(label = ind_num))+
  theme_bw(base_size = 10)+
  ggtitle(expression("PCA of filtered log"[2]*"cpm"))+
  guides(size="none")
Warning: ggrepel: 16 unlabeled data points (too many overlaps). Consider
increasing max.overlaps

Version Author Date
a07d5bc emmapfort 2025-03-19

Now, let’s make some correlation heatmaps These are pairwise across all genes by condition


#correlation heatmap for all samples

####PEARSON FILTERED####
fC_Matrix_Full_cpm_filter_pearsoncor <-
  cor(
    fC_Matrix_Full_cpm_filter,
    y = NULL,
    use = "everything",
    method = "pearson"
  )

####SPEARMAN FILTERED####
fC_Matrix_Full_cpm_filter_spearmancor <-
  cor(
    fC_Matrix_Full_cpm_filter,
    y = NULL,
    use = "everything",
    method = "spearman"
  )



#Now let's graph these correlations

pheatmap(fC_Matrix_Full_cpm_filter_pearsoncor, border_color = "black", legend = TRUE, angle_col = 90, display_numbers = FALSE, number_color = "black", fontsize = 12, fontsize_number = 9)

Version Author Date
a07d5bc emmapfort 2025-03-19

pheatmap(fC_Matrix_Full_cpm_filter_spearmancor, border_color = "black", legend = TRUE, angle_col = 90, display_numbers = FALSE, number_color = "black", fontsize = 12, fontsize_number = 9)

Version Author Date
a07d5bc emmapfort 2025-03-19



#now annotate these heatmaps
#Factor 1 - Individual
Individual <- as.factor(c(rep("Ind1", 9), rep("Ind2", 9), rep("Ind3", 9), rep("Ind4", 9), rep("Ind5", 9), rep("Ind6", 9), rep("Ind6REP", 9)))

#Factor 2 - Treatment
tx_factor <- c("DOX", "FLUO", "DMSO")
Tx <- as.factor(c(rep(tx_factor, 21)))
#view(Treatment)

#Factor 3 - Timepoint
time_factor <- c(rep("24", 3), rep("24rec", 3), rep("144rec", 3))
Time <- as.factor(c(rep(time_factor, 7)))

####annotation for colors####
annot_col_hm = list(Tx = c(DOX = "red", FLUO = "blue", DMSO = "black"),
                             Ind = c(Ind1 = "#66C2A5", Ind2 = "#FC8D62", Ind3 = "#1F78B4", Ind4 = "#E78AC3", Ind5 = "#A6D854", Ind6 = "#FFD92A", Ind6REP = "#8B3E9B"),
                             Time = c("24" = "#046A38", "24rec" = "#0050B5", "144rec" = "#B725AD"))

####annotation for values####
annot_list_hm <- data.frame(Individual = as.factor(c(rep("Ind1", 9), rep("Ind2", 9), rep("Ind3", 9), rep("Ind4", 9), rep("Ind5", 9), rep("Ind6", 9), rep("Ind6REP", 9))),
                                               Tx = as.factor(c(rep(tx_factor, 21))), 
                                               Time = as.factor(c(rep(time_factor, 7))))

  
##add in the annotations from above into the dataframe
row.names(annot_list_hm) <- colnames(fC_Matrix_Full_cpm_filter_pearsoncor)
row.names(annot_list_hm) <- colnames(fC_Matrix_Full_cpm_filter_spearmancor)

####ANNOTATED HEATMAPS####
pheatmap(fC_Matrix_Full_cpm_filter_pearsoncor, border_color = "black", legend = TRUE, angle_col = 90, display_numbers = FALSE, number_color = "black", fontsize = 10, fontsize_number = 5, annotation_col = annot_list_hm, annotation_colors = annot_col_hm)

Version Author Date
a07d5bc emmapfort 2025-03-19

pheatmap(fC_Matrix_Full_cpm_filter_spearmancor, border_color = "black", legend = TRUE, angle_col = 90, display_numbers = FALSE, number_color = "black", fontsize = 10, fontsize_number = 5, annotation_col = annot_list_hm, annotation_colors = annot_col_hm)

Version Author Date
a07d5bc emmapfort 2025-03-19

sessionInfo()
R version 4.4.2 (2024-10-31 ucrt)
Platform: x86_64-w64-mingw32/x64
Running under: Windows 11 x64 (build 22000)

Matrix products: default


locale:
[1] LC_COLLATE=English_United States.utf8 
[2] LC_CTYPE=English_United States.utf8   
[3] LC_MONETARY=English_United States.utf8
[4] LC_NUMERIC=C                          
[5] LC_TIME=English_United States.utf8    

time zone: America/Chicago
tzcode source: internal

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods  
[8] base     

other attached packages:
 [1] Rfast_2.1.5.1               RcppParallel_5.1.10        
 [3] zigg_0.0.2                  Rcpp_1.0.13-1              
 [5] RUVSeq_1.40.0               EDASeq_2.40.0              
 [7] ShortRead_1.64.0            GenomicAlignments_1.42.0   
 [9] SummarizedExperiment_1.36.0 MatrixGenerics_1.18.1      
[11] matrixStats_1.4.1           Rsamtools_2.22.0           
[13] GenomicRanges_1.58.0        Biostrings_2.74.0          
[15] GenomeInfoDb_1.42.3         XVector_0.46.0             
[17] IRanges_2.40.0              S4Vectors_0.44.0           
[19] BiocParallel_1.40.0         biomaRt_2.62.1             
[21] RColorBrewer_1.1-3          Cormotif_1.52.0            
[23] affy_1.84.0                 Biobase_2.66.0             
[25] BiocGenerics_0.52.0         PCAtools_2.18.0            
[27] ggfortify_0.4.17            pheatmap_1.0.12            
[29] edgeR_4.4.0                 limma_3.62.1               
[31] readxl_1.4.3                edgebundleR_0.1.4          
[33] lubridate_1.9.3             forcats_1.0.0              
[35] stringr_1.5.1               dplyr_1.1.4                
[37] purrr_1.0.2                 readr_2.1.5                
[39] tidyr_1.3.1                 tidyverse_2.0.0            
[41] tibble_3.2.1                hrbrthemes_0.8.7           
[43] reshape2_1.4.4              ggrepel_0.9.6              
[45] ggplot2_3.5.1               workflowr_1.7.1            

loaded via a namespace (and not attached):
  [1] later_1.4.1               BiocIO_1.16.0            
  [3] bitops_1.0-9              filelock_1.0.3           
  [5] R.oo_1.27.0               cellranger_1.1.0         
  [7] preprocessCore_1.68.0     XML_3.99-0.18            
  [9] lifecycle_1.0.4           httr2_1.1.0              
 [11] pwalign_1.2.0             rprojroot_2.0.4          
 [13] vroom_1.6.5               MASS_7.3-61              
 [15] processx_3.8.5            lattice_0.22-6           
 [17] magrittr_2.0.3            sass_0.4.9               
 [19] rmarkdown_2.29            jquerylib_0.1.4          
 [21] yaml_2.3.10               httpuv_1.6.15            
 [23] cowplot_1.1.3             DBI_1.2.3                
 [25] abind_1.4-8               zlibbioc_1.52.0          
 [27] R.utils_2.12.3            RCurl_1.98-1.16          
 [29] rappdirs_0.3.3            git2r_0.35.0             
 [31] gdtools_0.4.1             GenomeInfoDbData_1.2.13  
 [33] irlba_2.3.5.1             dqrng_0.4.1              
 [35] DelayedMatrixStats_1.28.1 codetools_0.2-20         
 [37] DelayedArray_0.32.0       xml2_1.3.6               
 [39] tidyselect_1.2.1          farver_2.1.2             
 [41] UCSC.utils_1.2.0          ScaledMatrix_1.14.0      
 [43] BiocFileCache_2.14.0      jsonlite_1.8.9           
 [45] systemfonts_1.1.0         tools_4.4.2              
 [47] progress_1.2.3            glue_1.8.0               
 [49] gridExtra_2.3             Rttf2pt1_1.3.12          
 [51] SparseArray_1.6.0         xfun_0.49                
 [53] withr_3.0.2               BiocManager_1.30.25      
 [55] fastmap_1.2.0             latticeExtra_0.6-30      
 [57] callr_3.7.6               digest_0.6.37            
 [59] rsvd_1.0.5                timechange_0.3.0         
 [61] R6_2.6.1                  mime_0.12                
 [63] colorspace_2.1-1          jpeg_0.1-10              
 [65] RSQLite_2.3.8             R.methodsS3_1.8.2        
 [67] generics_0.1.3            fontLiberation_0.1.0     
 [69] rtracklayer_1.66.0        prettyunits_1.2.0        
 [71] httr_1.4.7                htmlwidgets_1.6.4        
 [73] S4Arrays_1.6.0            whisker_0.4.1            
 [75] pkgconfig_2.0.3           gtable_0.3.6             
 [77] blob_1.2.4                hwriter_1.3.2.1          
 [79] htmltools_0.5.8.1         fontBitstreamVera_0.1.1  
 [81] scales_1.3.0              png_0.1-8                
 [83] knitr_1.49                rstudioapi_0.17.1        
 [85] tzdb_0.4.0                rjson_0.2.23             
 [87] curl_6.0.1                cachem_1.1.0             
 [89] parallel_4.4.2            extrafont_0.19           
 [91] AnnotationDbi_1.68.0      restfulr_0.0.15          
 [93] pillar_1.10.1             grid_4.4.2               
 [95] vctrs_0.6.5               promises_1.3.2           
 [97] BiocSingular_1.22.0       dbplyr_2.5.0             
 [99] beachmat_2.22.0           xtable_1.8-4             
[101] extrafontdb_1.0           evaluate_1.0.3           
[103] GenomicFeatures_1.58.0    cli_3.6.3                
[105] locfit_1.5-9.10           compiler_4.4.2           
[107] rlang_1.1.4               crayon_1.5.3             
[109] labeling_0.4.3            aroma.light_3.36.0       
[111] interp_1.1-6              ps_1.8.1                 
[113] getPass_0.2-4             plyr_1.8.9               
[115] fs_1.6.5                  stringi_1.8.4            
[117] deldir_2.0-4              munsell_0.5.1            
[119] fontquiver_0.2.1          Matrix_1.7-1             
[121] hms_1.1.3                 sparseMatrixStats_1.18.0 
[123] bit64_4.5.2               KEGGREST_1.46.0          
[125] statmod_1.5.0             shiny_1.10.0             
[127] igraph_2.1.1              memoise_2.0.1            
[129] affyio_1.76.0             bslib_0.9.0              
[131] bit_4.5.0