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library(GEOquery)
Loading required package: Biobase
Loading required package: BiocGenerics
Loading required package: parallel
Attaching package: 'BiocGenerics'
The following objects are masked from 'package:parallel':
clusterApply, clusterApplyLB, clusterCall, clusterEvalQ,
clusterExport, clusterMap, parApply, parCapply, parLapply,
parLapplyLB, parRapply, parSapply, parSapplyLB
The following objects are masked from 'package:stats':
IQR, mad, sd, var, xtabs
The following objects are masked from 'package:base':
anyDuplicated, append, as.data.frame, basename, cbind,
colMeans, colnames, colSums, dirname, do.call, duplicated,
eval, evalq, Filter, Find, get, grep, grepl, intersect,
is.unsorted, lapply, lengths, Map, mapply, match, mget, order,
paste, pmax, pmax.int, pmin, pmin.int, Position, rank, rbind,
Reduce, rowMeans, rownames, rowSums, sapply, setdiff, sort,
table, tapply, union, unique, unsplit, which, which.max,
which.min
Welcome to Bioconductor
Vignettes contain introductory material; view with
'browseVignettes()'. To cite Bioconductor, see
'citation("Biobase")', and for packages 'citation("pkgname")'.
Setting options('download.file.method.GEOquery'='auto')
Setting options('GEOquery.inmemory.gpl'=FALSE)
library(here)
here() starts at /nfsdata/projects/dylan/bentsen-rausch-2019
library(Seurat)
library(tidyverse)
── Attaching packages ────────────────────────────────── tidyverse 1.2.1 ──
✔ ggplot2 3.2.1 ✔ purrr 0.3.2
✔ tibble 2.1.3 ✔ dplyr 0.8.3
✔ tidyr 0.8.3 ✔ stringr 1.4.0
✔ readr 1.3.1.9000 ✔ forcats 0.4.0
── Conflicts ───────────────────────────────────── tidyverse_conflicts() ──
✖ dplyr::combine() masks Biobase::combine(), BiocGenerics::combine()
✖ dplyr::filter() masks stats::filter()
✖ dplyr::lag() masks stats::lag()
✖ ggplot2::Position() masks BiocGenerics::Position(), base::Position()
library(future)
library(data.table)
Attaching package: 'data.table'
The following objects are masked from 'package:dplyr':
between, first, last
The following object is masked from 'package:purrr':
transpose
library(rstatix)
Attaching package: 'rstatix'
The following object is masked from 'package:stats':
filter
plan(multiprocess, workers=30)
options(future.globals.maxSize = 8000 * 1024^2)
gse<-getGEO("GSE75330", GSEMatrix=T)
counts<-fread("/projects/dylan/fgf1/oligodendrocyte_figure/GSE75330_Marques_et_al_mol_counts2.tab")
counts<-as.data.frame(counts)
rownames(counts)<-counts[,1]
counts<-counts[,-1]
cellid<-pData(phenoData(gse[[1]]))[c(1,44)]
names(cellid)<-c("cell","label")
cellid<-data.frame(sapply(cellid, function(x) as.character(x)), stringsAsFactors = F)
counts<-counts[,colnames(counts)%in%cellid$cell]
cellid<-cellid[cellid$cell%in%colnames(counts),]
cellid<-cellid[match(colnames(counts),cellid$cell),]
CreateSeuratObject(counts=counts) %>%
SCTransform(verbose = F, return.only.var.genes = F) %>%
RunPCA(verbose=T) %>% RunUMAP(dims=1:10) -> branco
Warning: Feature names cannot have underscores ('_'), replacing with dashes
('-')
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Warning: The default method for RunUMAP has changed from calling Python UMAP via reticulate to the R-native UWOT using the cosine metric
To use Python UMAP via reticulate, set umap.method to 'umap-learn' and metric to 'correlation'
This message will be shown once per session
branco$label<-cellid$label
Idents(branco)<-"label"
branco_sub<-subset(branco, cells=WhichCells(branco, idents=c("PPR"), invert=T))
branco_sub<-RenameIdents(branco_sub, "MOL2"="MOL1")
branco_sub<-RenameIdents(branco_sub, "MOL3"="MOL1")
branco_sub<-RenameIdents(branco_sub, "MOL4"="MOL1")
branco_sub<-RenameIdents(branco_sub, "MOL5"="MOL1")
branco_sub<-RenameIdents(branco_sub, "MOL6"="MOL1")
branco_sub<-RenameIdents(branco_sub, "NFOL1"="NFOL")
branco_sub<-RenameIdents(branco_sub, "NFOL2"="NFOL")
branco_sub<-RenameIdents(branco_sub, "MFOL1"="MFOL")
branco_sub<-RenameIdents(branco_sub, "MFOL2"="MFOL")
oliglist <- list(branco=branco_sub, olig=olig)
olig.features <- SelectIntegrationFeatures(object.list = oliglist, nfeatures = 3000)
oliglist <- PrepSCTIntegration(object.list = oliglist, anchor.features = olig.features,
verbose = FALSE)
olig.anchors <- FindTransferAnchors(reference = oliglist[["branco"]],
query = oliglist[["olig"]], normalization.method = "SCT",
features = olig.features)
predictions <- TransferData(anchorset = olig.anchors, refdata = branco_sub@active.ident,
dims = 1:30)
hist(predictions$prediction.score.max)
Version | Author | Date |
---|---|---|
3b5cbe7 | Full Name | 2019-10-28 |
olig <- AddMetaData(object = olig, metadata = predictions)
table(olig$predicted.id)
COP MFOL MOL1 NFOL OPC
1004 2172 8049 760 3761
DimPlot(olig, group.by = "predicted.id")
Version | Author | Date |
---|---|---|
3b5cbe7 | Full Name | 2019-10-28 |
olig_sub<-subset(olig, subset=prediction.score.max>0.5)
olig_sub %>%
ScaleData(verbose = F, vars.to.regress = c("percent.mito","percent.ribo")) %>%
RunPCA(verbose=T) %>% FindNeighbors(dims = 1:20) %>% RunUMAP(dims = 1:20) %>%
FindClusters(resolution = 0.2) -> olig_sub
Modularity Optimizer version 1.3.0 by Ludo Waltman and Nees Jan van Eck
Number of nodes: 15274
Number of edges: 509268
Running Louvain algorithm...
Maximum modularity in 10 random starts: 0.9281
Number of communities: 8
Elapsed time: 3 seconds
DimPlot(olig_sub, label = T)
Warning: Using `as.character()` on a quosure is deprecated as of rlang 0.3.0.
Please use `as_label()` or `as_name()` instead.
This warning is displayed once per session.
Version | Author | Date |
---|---|---|
3b5cbe7 | Full Name | 2019-10-28 |
olig_sub <- subset(olig_sub, ident=c(3,4,6), invert=T)
olig_sub %>%
ScaleData(verbose = F, vars.to.regress = c("percent.mito","percent.ribo")) %>%
RunPCA(verbose=T) %>% FindNeighbors(dims = 1:20) %>% RunUMAP(dims = 1:20) %>%
FindClusters(resolution = 0.2) -> olig_sub
Warning in PrepDR(object = object, features = features, verbose = verbose):
The following 1 features requested have zero variance (running reduction
without them): 9130410C08Rik
Modularity Optimizer version 1.3.0 by Ludo Waltman and Nees Jan van Eck
Number of nodes: 13543
Number of edges: 432866
Running Louvain algorithm...
Maximum modularity in 10 random starts: 0.9161
Number of communities: 5
Elapsed time: 2 seconds
Idents(olig_sub)<-"predicted.id"
DimPlot(olig_sub, label = T)
Version | Author | Date |
---|---|---|
3b5cbe7 | Full Name | 2019-10-28 |
sessionInfo()
R version 3.5.3 (2019-03-11)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Storage
Matrix products: default
BLAS/LAPACK: /usr/lib64/libopenblas-r0.3.3.so
locale:
[1] LC_CTYPE=en_DK.UTF-8 LC_NUMERIC=C
[3] LC_TIME=en_DK.UTF-8 LC_COLLATE=en_DK.UTF-8
[5] LC_MONETARY=en_DK.UTF-8 LC_MESSAGES=en_DK.UTF-8
[7] LC_PAPER=en_DK.UTF-8 LC_NAME=C
[9] LC_ADDRESS=C LC_TELEPHONE=C
[11] LC_MEASUREMENT=en_DK.UTF-8 LC_IDENTIFICATION=C
attached base packages:
[1] parallel stats graphics grDevices utils datasets methods
[8] base
other attached packages:
[1] rstatix_0.1.1 data.table_1.12.2 future_1.14.0
[4] forcats_0.4.0 stringr_1.4.0 dplyr_0.8.3
[7] purrr_0.3.2 readr_1.3.1.9000 tidyr_0.8.3
[10] tibble_2.1.3 ggplot2_3.2.1 tidyverse_1.2.1
[13] Seurat_3.0.3.9036 here_0.1 GEOquery_2.50.5
[16] Biobase_2.42.0 BiocGenerics_0.28.0
loaded via a namespace (and not attached):
[1] Rtsne_0.15 colorspace_1.4-1 rio_0.5.16
[4] ggridges_0.5.1 rprojroot_1.3-2 fs_1.3.1
[7] rstudioapi_0.10 leiden_0.3.1 listenv_0.7.0
[10] npsurv_0.4-0 ggrepel_0.8.0.9000 RSpectra_0.15-0
[13] lubridate_1.7.4 xml2_1.2.0 codetools_0.2-16
[16] splines_3.5.3 R.methodsS3_1.7.1 lsei_1.2-0
[19] knitr_1.23 zeallot_0.1.0 jsonlite_1.6
[22] workflowr_1.4.0 broom_0.5.2 ica_1.0-2
[25] cluster_2.1.0 png_0.1-7 R.oo_1.22.0
[28] uwot_0.1.3 sctransform_0.2.0 compiler_3.5.3
[31] httr_1.4.1 backports_1.1.4 assertthat_0.2.1
[34] Matrix_1.2-17 lazyeval_0.2.2 cli_1.1.0
[37] limma_3.38.3 htmltools_0.3.6 tools_3.5.3
[40] rsvd_1.0.2 igraph_1.2.4.1 gtable_0.3.0
[43] glue_1.3.1 RANN_2.6.1 reshape2_1.4.3
[46] Rcpp_1.0.2 carData_3.0-2 cellranger_1.1.0
[49] vctrs_0.2.0 gdata_2.18.0 ape_5.3
[52] nlme_3.1-140 gbRd_0.4-11 lmtest_0.9-37
[55] xfun_0.8 globals_0.12.4 openxlsx_4.1.0.1
[58] rvest_0.3.4 irlba_2.3.3 gtools_3.8.1
[61] MASS_7.3-51.4 zoo_1.8-6 scales_1.0.0
[64] hms_0.5.0 RColorBrewer_1.1-2 curl_4.0
[67] yaml_2.2.0 reticulate_1.13 pbapply_1.4-1
[70] gridExtra_2.3 stringi_1.4.3 highr_0.8
[73] caTools_1.17.1.2 zip_2.0.3 bibtex_0.4.2
[76] Rdpack_0.11-0 SDMTools_1.1-221.1 rlang_0.4.0
[79] pkgconfig_2.0.2 bitops_1.0-6 evaluate_0.14
[82] lattice_0.20-38 ROCR_1.0-7 labeling_0.3
[85] htmlwidgets_1.3 cowplot_1.0.0 tidyselect_0.2.5
[88] RcppAnnoy_0.0.12 plyr_1.8.4 magrittr_1.5
[91] R6_2.4.0 gplots_3.0.1.1 generics_0.0.2
[94] foreign_0.8-71 withr_2.1.2 haven_2.1.0
[97] pillar_1.4.2 whisker_0.3-2 fitdistrplus_1.0-14
[100] abind_1.4-5 survival_2.44-1.1 future.apply_1.3.0
[103] tsne_0.1-3 car_3.0-3 modelr_0.1.4
[106] crayon_1.3.4 KernSmooth_2.23-15 plotly_4.9.0
[109] rmarkdown_1.13 readxl_1.3.1 grid_3.5.3
[112] git2r_0.25.2 metap_1.1 digest_0.6.20
[115] R.utils_2.9.0 RcppParallel_4.4.3 munsell_0.5.0
[118] viridisLite_0.3.0