Mediation analysis double filtered

Last updated: 2020-01-22

Checks: 7 0

Knit directory: Comparative_APA/analysis/

This reproducible R Markdown analysis was created with workflowr (version 1.5.0). The Checks tab describes the reproducibility checks that were applied when the results were created. The Past versions tab lists the development history.

R Markdown file: up-to-date

Great! Since the R Markdown file has been committed to the Git repository, you know the exact version of the code that produced these results.

Environment: empty

Great job! The global environment was empty. Objects defined in the global environment can affect the analysis in your R Markdown file in unknown ways. For reproduciblity it’s best to always run the code in an empty environment.

Seed: set.seed(20190902)

The command set.seed(20190902) was run prior to running the code in the R Markdown file. Setting a seed ensures that any results that rely on randomness, e.g. subsampling or permutations, are reproducible.

Session information: recorded

Great job! Recording the operating system, R version, and package versions is critical for reproducibility.

Cache: none

Nice! There were no cached chunks for this analysis, so you can be confident that you successfully produced the results during this run.

File paths: relative

Great job! Using relative paths to the files within your workflowr project makes it easier to run your code on other machines.

Repository version: 97ef45e

Great! You are using Git for version control. Tracking code development and connecting the code version to the results is critical for reproducibility. The version displayed above was the version of the Git repository at the time these results were generated.

Note that you need to be careful to ensure that all relevant files for the analysis have been committed to Git prior to generating the results (you can use wflow_publish or wflow_git_commit). workflowr only checks the R Markdown file, but you know if there are other scripts or data files that it depends on. Below is the status of the Git repository when the results were generated:


Ignored files:
    Ignored:    .DS_Store
    Ignored:    .Rhistory
    Ignored:    .Rproj.user/
    Ignored:    code/chimp_log/
    Ignored:    code/human_log/
    Ignored:    data/.DS_Store
    Ignored:    data/metadata_HCpanel.txt.sb-a5794dd2-i594qs/

Untracked files:
    Untracked:  ._.DS_Store
    Untracked:  Chimp/
    Untracked:  Human/
    Untracked:  analysis/CrossChimpThreePrime.Rmd
    Untracked:  analysis/DiffTransProtvsExpression.Rmd
    Untracked:  analysis/NormalizePheno_nuclear_DF.Rmd
    Untracked:  analysis/assessReadQual.Rmd
    Untracked:  analysis/diffExpressionPantro6.Rmd
    Untracked:  code/._ClassifyLeafviz.sh
    Untracked:  code/._Config_chimp.yaml
    Untracked:  code/._Config_chimp_full.yaml
    Untracked:  code/._Config_human.yaml
    Untracked:  code/._ConvertJunc2Bed.sh
    Untracked:  code/._CountNucleotides.py
    Untracked:  code/._CrossMapChimpRNA.sh
    Untracked:  code/._CrossMapThreeprime.sh
    Untracked:  code/._DiffSplice.sh
    Untracked:  code/._DiffSplicePlots.sh
    Untracked:  code/._DiffSplicePlots_gencode.sh
    Untracked:  code/._DiffSplice_gencode.sh
    Untracked:  code/._DiffSplice_removebad.sh
    Untracked:  code/._FindIntronForDomPAS.sh
    Untracked:  code/._FindIntronForDomPAS_DF.sh
    Untracked:  code/._GetMAPQscore.py
    Untracked:  code/._GetSecondaryMap.py
    Untracked:  code/._Lift5perPAS.sh
    Untracked:  code/._LiftFinalChimpJunc2Human.sh
    Untracked:  code/._LiftOrthoPAS2chimp.sh
    Untracked:  code/._MapBadSamples.sh
    Untracked:  code/._PAS_ATTAAA.sh
    Untracked:  code/._PAS_ATTAAA_df.sh
    Untracked:  code/._PASsequences.sh
    Untracked:  code/._PASsequences_DF.sh
    Untracked:  code/._PlotNuclearUsagebySpecies.R
    Untracked:  code/._PlotNuclearUsagebySpecies_DF.R
    Untracked:  code/._QuantMergedClusters.sh
    Untracked:  code/._ReverseLiftFilter.R
    Untracked:  code/._RunFixLeafCluster.sh
    Untracked:  code/._RunNegMCMediation.sh
    Untracked:  code/._RunNegMCMediationDF.sh
    Untracked:  code/._RunPosMCMediationDF.err
    Untracked:  code/._RunPosMCMediationDF.sh
    Untracked:  code/._Snakefile
    Untracked:  code/._SnakefilePAS
    Untracked:  code/._SnakefilePASfilt
    Untracked:  code/._SortIndexBadSamples.sh
    Untracked:  code/._bed215upbed.py
    Untracked:  code/._bed2SAF_gen.py
    Untracked:  code/._buildIndecpantro5
    Untracked:  code/._buildIndecpantro5.sh
    Untracked:  code/._buildLeafviz.sh
    Untracked:  code/._buildLeafviz_leadAnno.sh
    Untracked:  code/._buildStarIndex.sh
    Untracked:  code/._chimpChromprder.sh
    Untracked:  code/._cleanbed2saf.py
    Untracked:  code/._cluster.json
    Untracked:  code/._cluster2bed.py
    Untracked:  code/._clusterLiftReverse.sh
    Untracked:  code/._clusterLiftReverse_removebad.sh
    Untracked:  code/._clusterLiftprimary.sh
    Untracked:  code/._clusterLiftprimary_removebad.sh
    Untracked:  code/._converBam2Junc.sh
    Untracked:  code/._converBam2Junc_removeBad.sh
    Untracked:  code/._extraSnakefiltpas
    Untracked:  code/._filter5percPAS.py
    Untracked:  code/._filterNumChroms.py
    Untracked:  code/._filterPASforMP.py
    Untracked:  code/._filterPostLift.py
    Untracked:  code/._fixExonFC.py
    Untracked:  code/._fixLeafCluster.py
    Untracked:  code/._fixLiftedJunc.py
    Untracked:  code/._fixUTRexonanno.py
    Untracked:  code/._formathg38Anno.py
    Untracked:  code/._formatpantro6Anno.py
    Untracked:  code/._getRNAseqMapStats.sh
    Untracked:  code/._hg19MapStats.sh
    Untracked:  code/._humanChromorder.sh
    Untracked:  code/._intersectLiftedPAS.sh
    Untracked:  code/._liftJunctionFiles.sh
    Untracked:  code/._liftPAS19to38.sh
    Untracked:  code/._liftedchimpJunc2human.sh
    Untracked:  code/._makeNuclearDapaplots.sh
    Untracked:  code/._makeNuclearDapaplots_DF.sh
    Untracked:  code/._makeSamplyGroupsHuman_TvN.py
    Untracked:  code/._mapRNAseqhg19.sh
    Untracked:  code/._mapRNAseqhg19_newPipeline.sh
    Untracked:  code/._maphg19.sh
    Untracked:  code/._maphg19_subjunc.sh
    Untracked:  code/._mediation_test.R
    Untracked:  code/._mergeChimp3prime_inhg38.sh
    Untracked:  code/._mergedBam2BW.sh
    Untracked:  code/._nameClusters.py
    Untracked:  code/._negativeMediation_montecarlo.R
    Untracked:  code/._negativeMediation_montecarloDF.R
    Untracked:  code/._numMultimap.py
    Untracked:  code/._overlapapaQTLPAS.sh
    Untracked:  code/._postiveMediation_montecarlo_DF.R
    Untracked:  code/._prepareCleanLiftedFC_5perc4LC.py
    Untracked:  code/._prepareLeafvizAnno.sh
    Untracked:  code/._preparePAS4lift.py
    Untracked:  code/._primaryLift.sh
    Untracked:  code/._processhg38exons.py
    Untracked:  code/._quantJunc.sh
    Untracked:  code/._quantJunc_TEST.sh
    Untracked:  code/._quantJunc_removeBad.sh
    Untracked:  code/._quantMerged_seperatly.sh
    Untracked:  code/._recLiftchim2human.sh
    Untracked:  code/._revLiftPAShg38to19.sh
    Untracked:  code/._reverseLift.sh
    Untracked:  code/._runCheckReverseLift.sh
    Untracked:  code/._runChimpDiffIso.sh
    Untracked:  code/._runCountNucleotides.sh
    Untracked:  code/._runFilterNumChroms.sh
    Untracked:  code/._runHumanDiffIso.sh
    Untracked:  code/._runNuclearDiffIso_DF.sh
    Untracked:  code/._runNuclearDifffIso.sh
    Untracked:  code/._runTotalDiffIso.sh
    Untracked:  code/._run_chimpverifybam.sh
    Untracked:  code/._run_verifyBam.sh
    Untracked:  code/._snakemake.batch
    Untracked:  code/._snakemakePAS.batch
    Untracked:  code/._snakemakePASchimp.batch
    Untracked:  code/._snakemakePAShuman.batch
    Untracked:  code/._snakemake_chimp.batch
    Untracked:  code/._snakemake_human.batch
    Untracked:  code/._snakemakefiltPAS.batch
    Untracked:  code/._snakemakefiltPAS_chimp
    Untracked:  code/._snakemakefiltPAS_chimp.sh
    Untracked:  code/._snakemakefiltPAS_human.sh
    Untracked:  code/._submit-snakemake-chimp.sh
    Untracked:  code/._submit-snakemake-human.sh
    Untracked:  code/._submit-snakemakePAS-chimp.sh
    Untracked:  code/._submit-snakemakePAS-human.sh
    Untracked:  code/._submit-snakemakefiltPAS-chimp.sh
    Untracked:  code/._submit-snakemakefiltPAS-human.sh
    Untracked:  code/._subset_diffisopheno_Nuclear_HvC.py
    Untracked:  code/._subset_diffisopheno_Nuclear_HvC_DF.py
    Untracked:  code/._subset_diffisopheno_Total_HvC.py
    Untracked:  code/._threeprimeOrthoFC.sh
    Untracked:  code/._transcriptDTplotsNuclear.sh
    Untracked:  code/._verifyBam4973.sh
    Untracked:  code/._verifyBam4973inHuman.sh
    Untracked:  code/._wrap_chimpverifybam.sh
    Untracked:  code/._wrap_verifyBam.sh
    Untracked:  code/._writeMergecode.py
    Untracked:  code/.snakemake/
    Untracked:  code/ClassifyLeafviz.sh
    Untracked:  code/Config_chimp.yaml
    Untracked:  code/Config_chimp_full.yaml
    Untracked:  code/Config_human.yaml
    Untracked:  code/ConvertJunc2Bed.err
    Untracked:  code/ConvertJunc2Bed.out
    Untracked:  code/ConvertJunc2Bed.sh
    Untracked:  code/CountNucleotides.py
    Untracked:  code/CrossMapChimpRNA.sh
    Untracked:  code/CrossMapThreeprime.sh
    Untracked:  code/CrossmapChimp3prime.err
    Untracked:  code/CrossmapChimp3prime.out
    Untracked:  code/CrossmapChimpRNA.err
    Untracked:  code/CrossmapChimpRNA.out
    Untracked:  code/DiffSplice.err
    Untracked:  code/DiffSplice.out
    Untracked:  code/DiffSplice.sh
    Untracked:  code/DiffSplicePlots.err
    Untracked:  code/DiffSplicePlots.out
    Untracked:  code/DiffSplicePlots.sh
    Untracked:  code/DiffSplicePlots_gencode.sh
    Untracked:  code/DiffSplice_gencode.sh
    Untracked:  code/DiffSplice_removebad.err
    Untracked:  code/DiffSplice_removebad.out
    Untracked:  code/DiffSplice_removebad.sh
    Untracked:  code/FilterReverseLift.err
    Untracked:  code/FilterReverseLift.out
    Untracked:  code/FindIntronForDomPAS.err
    Untracked:  code/FindIntronForDomPAS.out
    Untracked:  code/FindIntronForDomPAS.sh
    Untracked:  code/FindIntronForDomPAS_DF.sh
    Untracked:  code/GencodeDiffSplice.err
    Untracked:  code/GencodeDiffSplice.out
    Untracked:  code/GetMAPQscore.py
    Untracked:  code/GetSecondaryMap.py
    Untracked:  code/HchromOrder.err
    Untracked:  code/HchromOrder.out
    Untracked:  code/JunctionLift.err
    Untracked:  code/JunctionLift.out
    Untracked:  code/JunctionLiftFinalChimp.err
    Untracked:  code/JunctionLiftFinalChimp.out
    Untracked:  code/Lift5perPAS.sh
    Untracked:  code/Lift5perPASbed.err
    Untracked:  code/Lift5perPASbed.out
    Untracked:  code/LiftClustersFirst.err
    Untracked:  code/LiftClustersFirst.out
    Untracked:  code/LiftClustersFirst_remove.err
    Untracked:  code/LiftClustersFirst_remove.out
    Untracked:  code/LiftClustersSecond.err
    Untracked:  code/LiftClustersSecond.out
    Untracked:  code/LiftClustersSecond_remove.err
    Untracked:  code/LiftClustersSecond_remove.out
    Untracked:  code/LiftFinalChimpJunc2Human.sh
    Untracked:  code/LiftOrthoPAS2chimp.sh
    Untracked:  code/LiftorthoPAS.err
    Untracked:  code/LiftorthoPASt.out
    Untracked:  code/Log.out
    Untracked:  code/MapBadSamples.err
    Untracked:  code/MapBadSamples.out
    Untracked:  code/MapBadSamples.sh
    Untracked:  code/MapStats.err
    Untracked:  code/MapStats.out
    Untracked:  code/MergeClusters.err
    Untracked:  code/MergeClusters.out
    Untracked:  code/MergeClusters.sh
    Untracked:  code/PAS_ATTAAA.err
    Untracked:  code/PAS_ATTAAA.out
    Untracked:  code/PAS_ATTAAA.sh
    Untracked:  code/PAS_ATTAAADF.err
    Untracked:  code/PAS_ATTAAADF.out
    Untracked:  code/PAS_ATTAAA_df.sh
    Untracked:  code/PAS_sequence.err
    Untracked:  code/PAS_sequence.out
    Untracked:  code/PAS_sequenceDF.err
    Untracked:  code/PAS_sequenceDF.out
    Untracked:  code/PASsequences.sh
    Untracked:  code/PASsequences_DF.sh
    Untracked:  code/PlotNuclearUsagebySpecies.R
    Untracked:  code/PlotNuclearUsagebySpecies_DF.R
    Untracked:  code/QuantMergeClusters
    Untracked:  code/QuantMergeClusters.err
    Untracked:  code/QuantMergeClusters.out
    Untracked:  code/QuantMergedClusters.sh
    Untracked:  code/Rev_liftoverPAShg19to38.err
    Untracked:  code/Rev_liftoverPAShg19to38.out
    Untracked:  code/ReverseLiftFilter.R
    Untracked:  code/RunFixCluster.err
    Untracked:  code/RunFixCluster.out
    Untracked:  code/RunFixLeafCluster.sh
    Untracked:  code/RunNegMCMediation.err
    Untracked:  code/RunNegMCMediation.sh
    Untracked:  code/RunNegMCMediationDF.err
    Untracked:  code/RunNegMCMediationDF.out
    Untracked:  code/RunNegMCMediationDF.sh
    Untracked:  code/RunNegMCMediationr.out
    Untracked:  code/RunPosMCMediation.err
    Untracked:  code/RunPosMCMediation.sh
    Untracked:  code/RunPosMCMediationDF.err
    Untracked:  code/RunPosMCMediationDF.out
    Untracked:  code/RunPosMCMediationDF.sh
    Untracked:  code/RunPosMCMediationr.out
    Untracked:  code/SAF215upbed_gen.py
    Untracked:  code/Snakefile
    Untracked:  code/SnakefilePAS
    Untracked:  code/SnakefilePASfilt
    Untracked:  code/SortIndexBadSamples.err
    Untracked:  code/SortIndexBadSamples.out
    Untracked:  code/SortIndexBadSamples.sh
    Untracked:  code/TotalTranscriptDTplot.err
    Untracked:  code/TotalTranscriptDTplot.out
    Untracked:  code/Upstream10Bases_general.py
    Untracked:  code/apaQTLsnake.err
    Untracked:  code/apaQTLsnake.out
    Untracked:  code/apaQTLsnakePAS.err
    Untracked:  code/apaQTLsnakePAS.out
    Untracked:  code/apaQTLsnakePAShuman.err
    Untracked:  code/bam2junc.err
    Untracked:  code/bam2junc.out
    Untracked:  code/bam2junc_remove.err
    Untracked:  code/bam2junc_remove.out
    Untracked:  code/bed215upbed.py
    Untracked:  code/bed2SAF_gen.py
    Untracked:  code/bed2saf.py
    Untracked:  code/bg_to_cov.py
    Untracked:  code/buildIndecpantro5
    Untracked:  code/buildIndecpantro5.sh
    Untracked:  code/buildLeafviz.err
    Untracked:  code/buildLeafviz.out
    Untracked:  code/buildLeafviz.sh
    Untracked:  code/buildLeafviz_leadAnno.sh
    Untracked:  code/buildLeafviz_leafanno.err
    Untracked:  code/buildLeafviz_leafanno.out
    Untracked:  code/buildStarIndex.sh
    Untracked:  code/callPeaksYL.py
    Untracked:  code/chimpChromprder.sh
    Untracked:  code/chooseAnno2Bed.py
    Untracked:  code/chooseAnno2SAF.py
    Untracked:  code/chromOrder.err
    Untracked:  code/chromOrder.out
    Untracked:  code/classifyLeafviz.err
    Untracked:  code/classifyLeafviz.out
    Untracked:  code/cleanbed2saf.py
    Untracked:  code/cluster.json
    Untracked:  code/cluster2bed.py
    Untracked:  code/clusterLiftReverse.sh
    Untracked:  code/clusterLiftReverse_removebad.sh
    Untracked:  code/clusterLiftprimary.sh
    Untracked:  code/clusterLiftprimary_removebad.sh
    Untracked:  code/clusterPAS.json
    Untracked:  code/clusterfiltPAS.json
    Untracked:  code/comands2Mege.sh
    Untracked:  code/converBam2Junc.sh
    Untracked:  code/converBam2Junc_removeBad.sh
    Untracked:  code/convertNumeric.py
    Untracked:  code/environment.yaml
    Untracked:  code/extraSnakefiltpas
    Untracked:  code/filter5perc.R
    Untracked:  code/filter5percPAS.py
    Untracked:  code/filter5percPheno.py
    Untracked:  code/filterBamforMP.pysam2_gen.py
    Untracked:  code/filterJuncChroms.err
    Untracked:  code/filterJuncChroms.out
    Untracked:  code/filterMissprimingInNuc10_gen.py
    Untracked:  code/filterNumChroms.py
    Untracked:  code/filterPASforMP.py
    Untracked:  code/filterPostLift.py
    Untracked:  code/filterSAFforMP_gen.py
    Untracked:  code/filterSortBedbyCleanedBed_gen.R
    Untracked:  code/filterpeaks.py
    Untracked:  code/fixExonFC.py
    Untracked:  code/fixFChead.py
    Untracked:  code/fixFChead_bothfrac.py
    Untracked:  code/fixLeafCluster.py
    Untracked:  code/fixLiftedJunc.py
    Untracked:  code/fixUTRexonanno.py
    Untracked:  code/formathg38Anno.py
    Untracked:  code/generateStarIndex.err
    Untracked:  code/generateStarIndex.out
    Untracked:  code/generateStarIndexHuman.err
    Untracked:  code/generateStarIndexHuman.out
    Untracked:  code/getRNAseqMapStats.sh
    Untracked:  code/hg19MapStats.err
    Untracked:  code/hg19MapStats.out
    Untracked:  code/hg19MapStats.sh
    Untracked:  code/humanChromorder.sh
    Untracked:  code/humanFiles
    Untracked:  code/intersectAnno.err
    Untracked:  code/intersectAnno.out
    Untracked:  code/intersectAnnoExt.err
    Untracked:  code/intersectAnnoExt.out
    Untracked:  code/intersectLiftedPAS.sh
    Untracked:  code/leafcutter_merge_regtools_redo.py
    Untracked:  code/liftJunctionFiles.sh
    Untracked:  code/liftPAS19to38.sh
    Untracked:  code/liftoverPAShg19to38.err
    Untracked:  code/liftoverPAShg19to38.out
    Untracked:  code/log/
    Untracked:  code/make5percPeakbed.py
    Untracked:  code/makeFileID.py
    Untracked:  code/makeNuclearDapaplots.sh
    Untracked:  code/makeNuclearDapaplots_DF.sh
    Untracked:  code/makeNuclearPlots.err
    Untracked:  code/makeNuclearPlots.out
    Untracked:  code/makeNuclearPlotsDF.err
    Untracked:  code/makeNuclearPlotsDF.out
    Untracked:  code/makePheno.py
    Untracked:  code/makeSamplyGroupsChimp_TvN.py
    Untracked:  code/makeSamplyGroupsHuman_TvN.py
    Untracked:  code/mapRNAseqhg19.sh
    Untracked:  code/mapRNAseqhg19_newPipeline.sh
    Untracked:  code/maphg19.err
    Untracked:  code/maphg19.out
    Untracked:  code/maphg19.sh
    Untracked:  code/maphg19_new.err
    Untracked:  code/maphg19_new.out
    Untracked:  code/maphg19_sub.err
    Untracked:  code/maphg19_sub.out
    Untracked:  code/maphg19_subjunc.sh
    Untracked:  code/mediation_test.R
    Untracked:  code/merge.err
    Untracked:  code/mergeChimp3prime_inhg38.sh
    Untracked:  code/merge_leafcutter_clusters_redo.py
    Untracked:  code/mergeandsort_ChimpinHuman.err
    Untracked:  code/mergeandsort_ChimpinHuman.out
    Untracked:  code/mergedBam2BW.sh
    Untracked:  code/mergedbam2bw.err
    Untracked:  code/mergedbam2bw.out
    Untracked:  code/nameClusters.py
    Untracked:  code/namePeaks.py
    Untracked:  code/negativeMediation_montecarlo.R
    Untracked:  code/negativeMediation_montecarloDF.R
    Untracked:  code/nuclearTranscriptDTplot.err
    Untracked:  code/nuclearTranscriptDTplot.out
    Untracked:  code/numMultimap.py
    Untracked:  code/overlapPAS.err
    Untracked:  code/overlapPAS.out
    Untracked:  code/overlapapaQTLPAS.sh
    Untracked:  code/overlapapaQTLPAS_extended.sh
    Untracked:  code/overlapapaQTLPAS_samples.sh
    Untracked:  code/peak2PAS.py
    Untracked:  code/pheno2countonly.R
    Untracked:  code/postiveMediation_montecarlo.R
    Untracked:  code/postiveMediation_montecarlo_DF.R
    Untracked:  code/prepareAnnoLeafviz.err
    Untracked:  code/prepareAnnoLeafviz.out
    Untracked:  code/prepareCleanLiftedFC_5perc4LC.py
    Untracked:  code/prepareLeafvizAnno.sh
    Untracked:  code/preparePAS4lift.py
    Untracked:  code/prepare_phenotype_table.py
    Untracked:  code/primaryLift.err
    Untracked:  code/primaryLift.out
    Untracked:  code/primaryLift.sh
    Untracked:  code/processhg38exons.py
    Untracked:  code/quantJunc.sh
    Untracked:  code/quantJunc_TEST.sh
    Untracked:  code/quantJunc_removeBad.sh
    Untracked:  code/quantLiftedPAS.err
    Untracked:  code/quantLiftedPAS.out
    Untracked:  code/quantLiftedPAS.sh
    Untracked:  code/quatJunc.err
    Untracked:  code/quatJunc.out
    Untracked:  code/recChimpback2Human.err
    Untracked:  code/recChimpback2Human.out
    Untracked:  code/recLiftchim2human.sh
    Untracked:  code/revLift.err
    Untracked:  code/revLift.out
    Untracked:  code/revLiftPAShg38to19.sh
    Untracked:  code/reverseLift.sh
    Untracked:  code/runCheckReverseLift.sh
    Untracked:  code/runChimpDiffIso.sh
    Untracked:  code/runCountNucleotides.err
    Untracked:  code/runCountNucleotides.out
    Untracked:  code/runCountNucleotides.sh
    Untracked:  code/runCountNucleotidesPantro6.err
    Untracked:  code/runCountNucleotidesPantro6.out
    Untracked:  code/runCountNucleotides_pantro6.sh
    Untracked:  code/runFilterNumChroms.sh
    Untracked:  code/runHumanDiffIso.sh
    Untracked:  code/runNuclearDiffIso_DF.sh
    Untracked:  code/runNuclearDifffIso.sh
    Untracked:  code/runTotalDiffIso.sh
    Untracked:  code/run_Chimpleafcutter_ds.err
    Untracked:  code/run_Chimpleafcutter_ds.out
    Untracked:  code/run_Chimpverifybam.err
    Untracked:  code/run_Chimpverifybam.out
    Untracked:  code/run_Humanleafcutter_ds.err
    Untracked:  code/run_Humanleafcutter_ds.out
    Untracked:  code/run_Nuclearleafcutter_ds.err
    Untracked:  code/run_Nuclearleafcutter_ds.out
    Untracked:  code/run_Nuclearleafcutter_dsDF.err
    Untracked:  code/run_Nuclearleafcutter_dsDF.out
    Untracked:  code/run_Totalleafcutter_ds.err
    Untracked:  code/run_Totalleafcutter_ds.out
    Untracked:  code/run_chimpverifybam.sh
    Untracked:  code/run_verifyBam.sh
    Untracked:  code/run_verifybam.err
    Untracked:  code/run_verifybam.out
    Untracked:  code/slurm-62824013.out
    Untracked:  code/slurm-62825841.out
    Untracked:  code/slurm-62826116.out
    Untracked:  code/slurm-64108209.out
    Untracked:  code/slurm-64108521.out
    Untracked:  code/slurm-64108557.out
    Untracked:  code/snakePASChimp.err
    Untracked:  code/snakePASChimp.out
    Untracked:  code/snakePAShuman.out
    Untracked:  code/snakemake.batch
    Untracked:  code/snakemakeChimp.err
    Untracked:  code/snakemakeChimp.out
    Untracked:  code/snakemakeHuman.err
    Untracked:  code/snakemakeHuman.out
    Untracked:  code/snakemakePAS.batch
    Untracked:  code/snakemakePASFiltChimp.err
    Untracked:  code/snakemakePASFiltChimp.out
    Untracked:  code/snakemakePASFiltHuman.err
    Untracked:  code/snakemakePASFiltHuman.out
    Untracked:  code/snakemakePASchimp.batch
    Untracked:  code/snakemakePAShuman.batch
    Untracked:  code/snakemake_chimp.batch
    Untracked:  code/snakemake_human.batch
    Untracked:  code/snakemakefiltPAS.batch
    Untracked:  code/snakemakefiltPAS_chimp.sh
    Untracked:  code/snakemakefiltPAS_human.sh
    Untracked:  code/submit-snakemake-chimp.sh
    Untracked:  code/submit-snakemake-human.sh
    Untracked:  code/submit-snakemakePAS-chimp.sh
    Untracked:  code/submit-snakemakePAS-human.sh
    Untracked:  code/submit-snakemakefiltPAS-chimp.sh
    Untracked:  code/submit-snakemakefiltPAS-human.sh
    Untracked:  code/subset_diffisopheno.py
    Untracked:  code/subset_diffisopheno_Chimp_tvN.py
    Untracked:  code/subset_diffisopheno_Huma_tvN.py
    Untracked:  code/subset_diffisopheno_Nuclear_HvC.py
    Untracked:  code/subset_diffisopheno_Nuclear_HvC_DF.py
    Untracked:  code/subset_diffisopheno_Total_HvC.py
    Untracked:  code/test
    Untracked:  code/threeprimeOrthoFC.out
    Untracked:  code/threeprimeOrthoFC.sh
    Untracked:  code/threeprimeOrthoFCcd.err
    Untracked:  code/transcriptDTplotsNuclear.sh
    Untracked:  code/transcriptDTplotsTotal.sh
    Untracked:  code/verifyBam4973.sh
    Untracked:  code/verifyBam4973inHuman.sh
    Untracked:  code/verifybam4973.err
    Untracked:  code/verifybam4973.out
    Untracked:  code/verifybam4973HumanMap.err
    Untracked:  code/verifybam4973HumanMap.out
    Untracked:  code/wrap_Chimpverifybam.err
    Untracked:  code/wrap_Chimpverifybam.out
    Untracked:  code/wrap_chimpverifybam.sh
    Untracked:  code/wrap_verifyBam.sh
    Untracked:  code/wrap_verifybam.err
    Untracked:  code/wrap_verifybam.out
    Untracked:  code/writeMergecode.py
    Untracked:  data/._.DS_Store
    Untracked:  data/._HC_filenames.txt
    Untracked:  data/._HC_filenames.txt.sb-4426323c-IKIs0S
    Untracked:  data/._HC_filenames.xlsx
    Untracked:  data/._MapPantro6_meta.txt
    Untracked:  data/._MapPantro6_meta.txt.sb-a5794dd2-Cskmlm
    Untracked:  data/._MapPantro6_meta.xlsx
    Untracked:  data/._OppositeSpeciesMap.txt
    Untracked:  data/._OppositeSpeciesMap.txt.sb-a5794dd2-mayWJf
    Untracked:  data/._OppositeSpeciesMap.xlsx
    Untracked:  data/._RNASEQ_metadata.txt
    Untracked:  data/._RNASEQ_metadata.txt.sb-4426323c-TE4ns3
    Untracked:  data/._RNASEQ_metadata.txt.sb-51f67ae1-HXp7Gq
    Untracked:  data/._RNASEQ_metadata_2Removed.txt
    Untracked:  data/._RNASEQ_metadata_2Removed.txt.sb-4426323c-a4lBwx
    Untracked:  data/._RNASEQ_metadata_2Removed.xlsx
    Untracked:  data/._RNASEQ_metadata_stranded.txt
    Untracked:  data/._RNASEQ_metadata_stranded.txt.sb-a5794dd2-D659m2
    Untracked:  data/._RNASEQ_metadata_stranded.txt.sb-a5794dd2-ImNMoY
    Untracked:  data/._RNASEQ_metadata_stranded.txt.sb-e4bf31f0-ZGnGgl
    Untracked:  data/._RNASEQ_metadata_stranded.xlsx
    Untracked:  data/._metadata_HCpanel.txt
    Untracked:  data/._metadata_HCpanel.txt.sb-a3d92a2d-b9cYoF
    Untracked:  data/._metadata_HCpanel.txt.sb-a5794dd2-i594qs
    Untracked:  data/._metadata_HCpanel.txt.sb-f4823d1e-qihGek
    Untracked:  data/._metadata_HCpanel.xlsx
    Untracked:  data/._metadata_HCpanel_frompantro5.xlsx
    Untracked:  data/._~$RNASEQ_metadata.xlsx
    Untracked:  data/._~$metadata_HCpanel.xlsx
    Untracked:  data/._.xlsx
    Untracked:  data/CompapaQTLpas/
    Untracked:  data/DTmatrix/
    Untracked:  data/DiffExpression/
    Untracked:  data/DiffIso_Nuclear/
    Untracked:  data/DiffIso_Nuclear_DF/
    Untracked:  data/DiffIso_Total/
    Untracked:  data/DiffSplice/
    Untracked:  data/DiffSplice_liftedJunc/
    Untracked:  data/DiffSplice_removeBad/
    Untracked:  data/DominantPAS/
    Untracked:  data/DominantPAS_DF/
    Untracked:  data/EvalPantro5/
    Untracked:  data/HC_filenames.txt
    Untracked:  data/HC_filenames.xlsx
    Untracked:  data/Khan_prot/
    Untracked:  data/Li_eqtls/
    Untracked:  data/MapPantro6_meta.txt
    Untracked:  data/MapPantro6_meta.xlsx
    Untracked:  data/MapStats/
    Untracked:  data/NormalizedClusters/
    Untracked:  data/NuclearHvC/
    Untracked:  data/NuclearHvC_DF/
    Untracked:  data/OppositeSpeciesMap.txt
    Untracked:  data/OppositeSpeciesMap.xlsx
    Untracked:  data/OverlapBenchmark/
    Untracked:  data/PAS/
    Untracked:  data/PAS_doubleFilter/
    Untracked:  data/Peaks_5perc/
    Untracked:  data/Pheno_5perc/
    Untracked:  data/Pheno_5perc_DF_nuclear/
    Untracked:  data/Pheno_5perc_nuclear/
    Untracked:  data/Pheno_5perc_nuclear_old/
    Untracked:  data/Pheno_5perc_total/
    Untracked:  data/RNASEQ_metadata.txt
    Untracked:  data/RNASEQ_metadata_2Removed.txt
    Untracked:  data/RNASEQ_metadata_2Removed.xlsx
    Untracked:  data/RNASEQ_metadata_stranded.txt
    Untracked:  data/RNASEQ_metadata_stranded.txt.sb-e4bf31f0-ZGnGgl/
    Untracked:  data/RNASEQ_metadata_stranded.xlsx
    Untracked:  data/SignalSites/
    Untracked:  data/SignalSites_doublefilter/
    Untracked:  data/Threeprime2Ortho/
    Untracked:  data/TotalHvC/
    Untracked:  data/TwoBadSampleAnalysis/
    Untracked:  data/Wang_ribo/
    Untracked:  data/apaQTLGenes/
    Untracked:  data/chainFiles/
    Untracked:  data/cleanPeaks_anno/
    Untracked:  data/cleanPeaks_byspecies/
    Untracked:  data/cleanPeaks_lifted/
    Untracked:  data/files4viz_nuclear/
    Untracked:  data/files4viz_nuclear_DF/
    Untracked:  data/leafviz/
    Untracked:  data/liftover_files/
    Untracked:  data/mediation/
    Untracked:  data/mediation_DF/
    Untracked:  data/metadata_HCpanel.txt
    Untracked:  data/metadata_HCpanel.xlsx
    Untracked:  data/metadata_HCpanel_frompantro5.txt
    Untracked:  data/metadata_HCpanel_frompantro5.xlsx
    Untracked:  data/primaryLift/
    Untracked:  data/reverseLift/
    Untracked:  data/~$RNASEQ_metadata.xlsx
    Untracked:  data/~$metadata_HCpanel.xlsx
    Untracked:  data/.xlsx
    Untracked:  output/dtPlots/
    Untracked:  projectNotes.Rmd

Unstaged changes:
    Modified:   analysis/ExploredAPA.Rmd
    Modified:   analysis/OppositeMap.Rmd
    Modified:   analysis/annotationInfo.Rmd
    Modified:   analysis/comp2apaQTLPAS.Rmd
    Modified:   analysis/correlationPhenos.Rmd
    Modified:   analysis/establishCutoffs.Rmd
    Modified:   analysis/investigatePantro5.Rmd
    Modified:   analysis/multiMap.Rmd
    Modified:   analysis/speciesSpecific.Rmd

Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes.

These are the previous versions of the R Markdown and HTML files. If you’ve configured a remote Git repository (see ?wflow_git_remote), click on the hyperlinks in the table below to view them.

File	Version	Author	Date	Message
Rmd	97ef45e	brimittleman	2020-01-22	add mediation DF

library(workflowr)

This is workflowr version 1.5.0
Run ?workflowr for help getting started

library(tidyverse)

── Attaching packages ──────────────────────────────────────────────────────────────────────────────────────────── tidyverse 1.2.1 ──

✔ ggplot2 3.1.1       ✔ purrr   0.3.2  
✔ tibble  2.1.1       ✔ dplyr   0.8.0.1
✔ tidyr   0.8.3       ✔ stringr 1.3.1  
✔ readr   1.3.1       ✔ forcats 0.3.0

── Conflicts ─────────────────────────────────────────────────────────────────────────────────────────────── tidyverse_conflicts() ──
✖ dplyr::filter() masks stats::filter()
✖ dplyr::lag()    masks stats::lag()

library(limma)
library(MASS)


Attaching package: 'MASS'

The following object is masked from 'package:dplyr':

    select

In this analysis, I will ask if differrecnes in APA or splicing are causal for differences in expression. I did this before the filter, this is post the double filter for expression and 10% cutoff.

Ittai’s version of the analysis: https://ittaieres.github.io/HiCiPSC/gene_expression.html#now,_get_the_appropriate_data_and_actually_run_the_mediation_analysis

To deal with multiple phenotypes per gene I will take the highest absolute effect size cluster or PAS for splicing and APA respectively.

mkdir ../data/mediation_DF

source("../code/mediation_test.R") #Obtain necessary functions

Now I need to pull in the expression, apa, and splicing data.
- expression= log2RPKM, and adju.P.val - normalized nuclear apa phenotype
- splicing normalized clusters

Expression and APA

#expression
nameID=read.table("../../genome_anotation_data/ensemble_to_genename.txt",sep="\t", header = T, stringsAsFactors = F) %>% dplyr::select(Gene_stable_ID,Gene.name)
ExpRes=read.table("../data/DiffExpression/DEtested_allres.txt", header = F, stringsAsFactors = F, col.names = c("Gene_stable_ID", "logFC", "AveExpr", "t", "P.Value", "adj.P.Val", "B")) %>% inner_join(nameID,by="Gene_stable_ID") %>% dplyr::select(Gene.name,adj.P.Val )
ExpNorm=read.table("../data/DiffExpression/NormalizedExpressionPassCutoff.txt", header = T, stringsAsFactors = F) %>% inner_join(nameID,by="Gene_stable_ID") %>% dplyr::select(-Gene_stable_ID)
#join indiv with adjust pval
Exp=ExpRes %>% inner_join(ExpNorm, by="Gene.name")%>% rename("gene"= Gene.name) %>% dplyr::select(gene, adj.P.Val, NA18498, NA18499, NA18502, NA18504, NA18510,NA18523,NA18358, NA3622,NA3659,NA4973,NAPT30,NAPT91)

#Chr    start   end ID  NA18498_N   NA18499_N   NA18502_N   NA18504_N   NA18510_N   NA18523_N   NA18358_N   NA3622_N    NA3659_N    NA4973_N    NApt30_N    NApt91_N
#apa:  
apaN= read.table("../data/Pheno_5perc_DF_nuclear/ALLPAS_postLift_LocParsed_bothSpecies_pheno_DoubleFilter_Nuclear.txt.gz.qqnorm_AllChrom", col.names = c("chr", "start", "end", "id","NA18498_APA", "NA18499_APA", "NA18502_APA", "NA18504_APA", "NA18510_APA","NA18523_APA", "NA18358_APA", "NA3622_APA","NA3659_APA", "NA4973_APA","NAPT30_APA","NAPT91_APA")) %>% separate(id, into=c("ch", "st", "en","id2"),sep=":") %>% separate(id2, into=c("gene", "strand","id3"),sep="_") %>% separate(id3, into=c("loc", "disc", "PAS"), sep="-") %>% dplyr::select(gene, PAS,contains("NA"))
PASMeta=read.table("../data/PAS_doubleFilter/PAS_10perc_either_HumanCoord_BothUsage_meta_doubleFilter.txt", header = T, stringsAsFactors = F) %>% dplyr::select(PAS, chr, start,end, gene)
apaRes= read.table("../data/DiffIso_Nuclear_DF/TN_diff_isoform_allChrom.txt_significance.txt",sep="\t" ,col.names = c('status','loglr','df','p','cluster','p.adjust'),stringsAsFactors = F) %>% filter(status=="Success") %>% separate(cluster, into=c("chr","gene"),sep=":")
apaPASres=read.table("../data/DiffIso_Nuclear_DF/TN_diff_isoform_allChrom.txt_effect_sizes.txt", stringsAsFactors = F, col.names=c('intron',  'logef' ,'Human', 'Chimp','deltaPAU')) %>% filter(intron != "intron") %>% separate(intron, into=c("chr","start", "end","gene"), sep=":") 
apaPASres$start=as.integer(apaPASres$start)
apaPASres$end=as.integer(apaPASres$end)
apaPASres$deltaPAU=as.numeric(apaPASres$deltaPAU)
apaPASres=apaPASres%>% inner_join(PASMeta,by=c("chr", "start", "end", "gene"))
#problem if there are 2 pas then the are opposite but same value - do with all one direction for 
apaPASres_topPos= apaPASres %>% group_by(gene) %>% top_n(1,abs(deltaPAU)) %>% top_n(1,deltaPAU)
apaPASres_topNeg= apaPASres %>% group_by(gene) %>% top_n(1,abs(deltaPAU)) %>% top_n(-1,deltaPAU)
#join with pvalue: 
apaPASres_topPos_pval=apaPASres_topPos %>%  inner_join(apaRes, by=c("gene"))  %>% dplyr::select( PAS, p.adjust)

Adding missing grouping variables: `gene`

apaPASres_topNeg_pval=apaPASres_topNeg %>%  inner_join(apaRes, by=c("gene")) %>% dplyr::select( PAS, p.adjust)

Adding missing grouping variables: `gene`

#looks like the naming convention is not consistent...
apaPASres_topPos_pval_normAPA=apaPASres_topPos_pval %>% inner_join(apaN,by=c("PAS","gene")) %>% dplyr::select(-PAS,-p.adjust)
apaPASres_topNeg_pval_normAPA=apaPASres_topNeg_pval %>% inner_join(apaN,by=c("PAS","gene"))%>% dplyr::select(-PAS,-p.adjust)


#create a dataframe with expression and apa
APA_posandExp=Exp %>% inner_join(apaPASres_topPos_pval_normAPA, by="gene")
APA_negandExp=Exp %>% inner_join(apaPASres_topNeg_pval_normAPA,by="gene")

First do the positive:

Seperate De and not De

is_de <- which(APA_posandExp$adj.P.Val < .05)
isnot_de <- which(APA_posandExp$adj.P.Val >= .05)

gvec <- APA_posandExp$genes
Exp_pos <- APA_posandExp %>% dplyr::select(-gene,-adj.P.Val, -contains("APA"))
APA_pos <- APA_posandExp %>% dplyr::select(contains("APA"))

# metadata label
species <- factor(c("H","H","H","H","H","H","C","C","C","C","C","C"))
batch <- factor(c("A", "A", "B", "A", "B", "B", "B", "B","A", "A","A","B"))

metadata <- data.frame(sample=names(Exp_pos),
                       species=species,
                       batch=batch)

Compute indirect effects:

fit_de_pos <- test_mediation(exprs = Exp_pos[is_de,], 
                         fixed_covariates = list(species=metadata$species,
                                                 batch=metadata$batch),
                         varying_covariate = APA_pos[is_de,])

fit_node_pos <- test_mediation(exprs = Exp_pos[isnot_de,], 
                           fixed_covariates = list(species=metadata$species,
                                                   batch=metadata$batch),
                           varying_covariate = APA_pos[isnot_de,])

#save as R data object:
save(fit_de_pos, fit_node_pos, is_de, isnot_de, Exp_pos, APA_pos, metadata, gvec, file = "../data/mediation_DF/positive_mediation.rda")

Montecarlo:

I will have to run this seperatly as an Rscript.


sbatch RunPosMCMediationDF.sh

Load results

mc_pos_de <- readRDS(file = "../data/mediation_DF/mc_de_postive.rds")
mc_pos_node <- readRDS(file = "../data/mediation_DF/mc_node_postive.rds")

##In DE genes
ngenes <- ncol(mc_pos_de)
ab <- fit_de_pos$alpha*fit_de_pos$beta
out <- sapply(1:ngenes, function(g) {
  x <- unlist(mc_pos_de[,g])
  q <- quantile(x, prob = c(.025, .975))
#  q <- quantile(x, prob = c(.005, .995))
  ifelse(0 > q[1] & 0 < q[2], F, T)
})
table(out)

out
FALSE  TRUE 
 2679    85

85/(85+2679) #

[1] 0.03075253

DEdat <- data.frame(bf=fit_de_pos$tau, af=fit_de_pos$tau_prime,significance=out)
DEdat$color <- ifelse(DEdat$significance==TRUE, "red", "black")
plot(x=DEdat$bf, y=DEdat$af, ylab="Effect Size After Controlling for APA", xlab="Effect Size Before Controlling for APA", main="Effect of APA on Expression Divergence in DE genes", pch=16, cex=0.6, xlim=c(-10, 10), ylim=c(-10,10),col=alpha(DEdat$color, 0.6))
legend("topleft", legend=c("95% CI Significant (n=85)", "95% CI Non significant (n=2679)"), col=c("red", "black"), pch=16:16, cex=0.8)
abline(0, 1)
abline(h=0)
abline(v=0)

Not DE:

##In non-DE genes
ngenes <- ncol(mc_pos_node)
ab <- fit_node_pos$alpha*fit_node_pos$beta
out <- sapply(1:ngenes, function(g) {
  x <- unlist(mc_pos_node[,g])
  q <- quantile(x, prob = c(.025, .975))
#  q <- quantile(x, prob = c(.005, .995))
  ifelse(0 > q[1] & 0 < q[2], F, T)
})
table(out)

out
FALSE  TRUE 
 4557   116

116/(116+4557)

[1] 0.02482345

#Visualization for non DE genes:
noDEdat <- data.frame(bf=fit_node_pos$tau, af=fit_node_pos$tau_prime,significance=out)
noDEdat$color <- ifelse(noDEdat$significance==TRUE, "red", "black")
plot(x=noDEdat$bf, y=noDEdat$af, ylab="Effect Size After Controlling for APA", xlab="Effect Size Before Controlling for APA", main="Effect of APA on Expression Divergence in non-DE genes", pch=16, cex=0.6, xlim=c(-10, 10), ylim=c(-10,10), adj=0.6,col=alpha(noDEdat$color, 0.6))
legend("topleft", legend=c("95% CI Significant (n=116)", "95% CI Non significant (n=4557)"), col=c("red", "black"), pch=16:16, cex=0.8)
abline(0, 1)
abline(h=0)
abline(v=0)

Negative: