diff --git a/bash.scripts/sc_TissueMapper-D27PrF.sh b/bash.scripts/sc_TissueMapper-D27PrF.sh deleted file mode 100644 index 70eee7fae548fa4877a8d03c4f458c2b0ab71c4a..0000000000000000000000000000000000000000 --- a/bash.scripts/sc_TissueMapper-D27PrF.sh +++ /dev/null @@ -1,13 +0,0 @@ -#!/bin/bash -#SBATCH --job-name R_FullAnalysis -#SBATCH -p 256GB,256GBv1,384GB -#SBATCH -N 1 -#SBATCH -t 7-0:0:0 -#SBATCH -o job_%j.out -#SBATCH -e job_%j.out -#SBATCH --mail-type ALL -#SBATCH --mail-user gervaise.henry@utsouthwestern.edu - -module load R/3.4.1-gccmkl - -Rscript ../r.scripts/sc-TissueMapper_RUN.D27.R \ No newline at end of file diff --git a/bash.scripts/sc_TissueMapper-DPrF3.sh b/bash.scripts/sc_TissueMapper-DPrF3.sh deleted file mode 100644 index 557111e99bedb0c1275c87347aa9abb1f8292d0c..0000000000000000000000000000000000000000 --- a/bash.scripts/sc_TissueMapper-DPrF3.sh +++ /dev/null @@ -1,15 +0,0 @@ -#!/bin/bash -#SBATCH --job-name R_FullAnalysis -#SBATCH -p 256GB,256GBv1,384GB -#SBATCH -N 1 -#SBATCH -t 7-0:0:0 -#SBATCH -o job_%j.out -#SBATCH -e job_%j.out -#SBATCH --mail-type ALL -#SBATCH --mail-user gervaise.henry@utsouthwestern.edu - -module load R/3.4.1-gccmkl - -Rscript ../r.scripts/sc-TissueMapper_RUN.D3.R -#Rscript ../r.scripts/sc-TissueMapper_RUN.D.diy.R -#Rscript ../r.scripts/sc-TissueMapper_RUN.D.pseudotime.R \ No newline at end of file diff --git a/r.scripts/sc-TissueMapper_RUN.D27.R b/r.scripts/sc-TissueMapper_RUN.D27.R deleted file mode 100644 index 252034b3db5d8886b201c86c8ac7d8b49a82fd5d..0000000000000000000000000000000000000000 --- a/r.scripts/sc-TissueMapper_RUN.D27.R +++ /dev/null @@ -1,288 +0,0 @@ -gc() -library(methods) -library(optparse) -library(Seurat) -library(readr) -library(fBasics) -library(pastecs) -library(qusage) - -source("../r.scripts/sc-TissueMapper.R") - -#Create folder structure -setwd("../") -if (!dir.exists("./analysis")){ - dir.create("./analysis") -} -if (!dir.exists("./analysis/qc")){ - dir.create("./analysis/qc") -} -if (!dir.exists("./analysis/qc/cc")){ - dir.create("./analysis/qc/cc") -} -if (!dir.exists("./analysis/tSNE")){ - dir.create("./analysis/tSNE") -} -if (!dir.exists("./analysis/tSNE/pre.stress")){ - dir.create("./analysis/tSNE/pre.stress") -} -if (!dir.exists("./analysis/pca")){ - dir.create("./analysis/pca") -} -if (!dir.exists("./analysis/pca/stress")){ - dir.create("./analysis/pca/stress") -} -if (!dir.exists("./analysis/violin")){ - dir.create("./analysis/violin") -} -if (!dir.exists("./analysis/violin/stress")){ - dir.create("./analysis/violin/stress") -} -if (!dir.exists("./analysis/table")){ - dir.create("./analysis/table") -} -if (!dir.exists("./analysis/tSNE/post.stress")){ - dir.create("./analysis/tSNE/post.stress") -} -if (!dir.exists("./analysis/cor")){ - dir.create("./analysis/cor") -} -if (!dir.exists("./analysis/tSNE/lin")){ - dir.create("./analysis/tSNE/lin") -} -if (!dir.exists("./analysis/tSNE/epi")){ - dir.create("./analysis/tSNE/epi") -} -if (!dir.exists("./analysis/tSNE/st")){ - dir.create("./analysis/tSNE/st") -} -if (!dir.exists("./analysis/tSNE/merge")){ - dir.create("./analysis/tSNE/merge") -} -if (!dir.exists("./analysis/pca/ne")){ - dir.create("./analysis/pca/ne") -} -if (!dir.exists("./analysis/tSNE/ne")){ - dir.create("./analysis/tSNE/ne") -} -if (!dir.exists("./analysis/violin/ne")){ - dir.create("./analysis/violin/ne") -} -if (!dir.exists("./analysis/tSNE/FINAL")){ - dir.create("./analysis/tSNE/FINAL") -} -if (!dir.exists("./analysis/deg")){ - dir.create("./analysis/deg") -} - -#Retrieve command-line options -option_list=list( - make_option("--p",action="store",default="D27PrF",type='character',help="Project Name"), - make_option("--g",action="store",default="ALL",type='character',help="Group To analyze"), - make_option("--lg",action="store",default=500,type='integer',help="Threshold for cells with minimum genes"), - make_option("--hg",action="store",default=2500,type='integer',help="Threshold for cells with maximum genes"), - make_option("--lm",action="store",default=0,type='numeric',help="Threshold for cells with minimum %mito genes"), - make_option("--hm",action="store",default=0.1,type='numeric',help="Threshold for cells with maximum %mito genes"), - make_option("--lx",action="store",default=0.0125,type='numeric',help="x low threshold for hvg selection"), - make_option("--hx",action="store",default=3.5,type='numeric',help="x high threshold for hvg selection"), - make_option("--ly",action="store",default=0.5,type='numeric',help="y low threshold for hvg selection"), - make_option("--cc",action="store",default=TRUE,type='logical',help="Scale cell cycle?"), - make_option("--pc",action="store",default=25,type='integer',help="Number of PCs to cacluate"), - make_option("--hpc",action="store",default=0.8,type='numeric',help="Max variance cutoff for PCs to use, pre-stress"), - make_option("--res.prestress",action="store",default=1,type='numeric',help="Resolution to cluster, pre-stress"), - make_option("--st",action="store",default="TRUE",type='logical',help="Remove stressed cells?"), - make_option("--stg",action="store",default="dws",type='character',help="Geneset to use for stress ID"), - make_option("--hpc.poststress",action="store",default=0.80,type='numeric',help="Max variance cutoff for PCs to use, post-stress"), - make_option("--res.poststress",action="store",default=0.5,type='numeric',help="Resolution to cluster, post-stress"), - make_option("--ds",action="store",default=0,type='integer',help="Number of cells to downsample"), - make_option("--hpc.epi",action="store",default=0.75,type='numeric',help="Max variance cutoff for PCs to use, Epi"), - make_option("--res.epi",action="store",default=0.2,type='numeric',help="Resolution to cluster, Epi"), - make_option("--hpc.st",action="store",default=0.75,type='numeric',help="Max variance cutoff for PCs to use, St"), - make_option("--res.st",action="store",default=0.2,type='numeric',help="Resolution to cluster, St") - ) -opt=parse_args(OptionParser(option_list=option_list)) -rm(option_list) -if (opt$lm==0){opt$lm=-Inf} - -sc10x <- scLoad(opt$p) - -sc10x <- scSubset(sc10x,"ALL",opt$g) - -if (opt$cc==TRUE){ - results <- scCellCycle(sc10x) - sc10x <- results[[1]] - genes.s <- results[[2]] - genes.g2m <- results[[3]] - rm(results) -} else { - genes.s="" - genes.g2m="" -} - -results <- scQC(sc10x,lg=opt$lg,hg=opt$hg,lm=opt$lm,hm=opt$hm) -sc10x <- results[[1]] -counts.cell.raw <- results[[2]] -counts.gene.raw <- results[[3]] -counts.cell.filtered <- results[[4]] -counts.gene.filtered <- results[[5]] -rm(results) - -results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="pre.stress") -sc10x <- results[[1]] -genes.hvg <- results[[2]] -pc.use <- results[[3]] -rm(results) - -sc10x <- scCluster(sc10x,pc.use=pc.use,res.use=opt$res.prestress,folder="pre.stress") - -if (opt$st==TRUE){ - results <- scStress(sc10x,stg=opt$stg,res.use=opt$res.prestress,pc.use=pc.use) - sc10x <- results[[1]] - counts.cell.filtered.stress <- results[[2]] - sc10x.Stress <- results[[3]] - rm(results) - - results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc.poststress,file="post.stress") - sc10x <- results[[1]] - genes.hvg.poststress <- results[[2]] - pc.use.poststress <- results[[3]] - rm(results) - - sc10x <- scCluster(sc10x,pc.use=pc.use.poststress,res.use=opt$res.poststress,folder="post.stress") -} - -gene.set1 <- read_delim("./genesets/genes.deg.St.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "St" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.Epi.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Epi" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -results <- scQuSAGE(sc10x,gs=gene.set,res.use=opt$res.poststress,ds=opt$ds,nm="Lin",folder="lin") -sc10x <- results[[1]] -results.cor.Lin <- results[[2]] -results.clust.Lin.id <- results[[3]] -rm(results) -rm(gene.set) - -sc10x.Epi <- scSubset(sc10x,i="Lin",g="Epi") -if (any(levels(sc10x@ident)=="Unknown")){ - sc10x.St <- scSubset(sc10x,i="Lin",g=c("St","Unknown")) -} else { - sc10x.St <- scSubset(sc10x,i="Lin",g="St") -} - -#results <- scPC(sc10x.Epi,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc.epi,file="Epi") -#sc10x.Epi <- results[[1]] -#genes.hvg.epi <- results[[2]] -#pc.use.epi <- results[[3]] -#rm(results) - -sc10x.Epi <- scCluster(sc10x.Epi,pc.use=pc.use.poststress,res.use=0.1,folder="epi") -sc10x.Epi <- scCluster(sc10x.Epi,pc.use=pc.use.poststress,res.use=0.5,folder="epi") -sc10x.Epi <- scCluster(sc10x.Epi,pc.use=pc.use.poststress,res.use=opt$res.epi,folder="epi") - -gene.set1 <- read_delim("./genesets/genes.deg.BE.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "BE" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.LE.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "LE" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.OE1.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "OE_SCGB" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.OE2.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "OE_KRT13" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -results <- scQuSAGE(sc10x.Epi,gs=gene.set,res.use=opt$res.epi,ds=opt$ds,nm="Epi.dws.sc",folder="epi") -sc10x.Epi <- results[[1]] -results.cor.Epi.dws.sc <- results[[2]] -results.clust.Epi.dws.id <- results[[3]] -rm(results) -rm(gene.set) - -#results <- scPC(sc10x.St,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc.st,file="St") -#sc10x.St <- results[[1]] -#genes.hvg.st <- results[[2]] -#pc.use.st <- results[[3]] -#rm(results) - -sc10x.St <- scCluster(sc10x.St,pc.use=pc.use.poststress,res.use=0.1,folder="st") -sc10x.St <- scCluster(sc10x.St,pc.use=pc.use.poststress,res.use=0.5,folder="st") -sc10x.St <- scCluster(sc10x.St,pc.use=pc.use.poststress,res.use=opt$res.st,folder="st") - -gene.set1 <- read_delim("./genesets/genes.deg.Fib.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Fib" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.SM.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "SM" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.Endo.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Endo" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.Leu.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Leu" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -results <- scQuSAGE(sc10x.St,gs=gene.set,res.use=opt$res.st,ds=opt$ds,nm="St.dws.sc",folder="st") -sc10x.St <- results[[1]] -results.cor.St.go <- results[[2]] -results.clust.St.go.id <- results[[3]] -rm(results) -rm(gene.set) - -sc10x.Epi.NE <- scNE(sc10x.Epi,neg="dws") - -sc10x.Epi <- scMergeSubClust(sc10x.Epi,i="Epi.dws.sc",g=c("BE","LE","OE_SCGB","OE_KRT13"),nm="Merge") - -sc10x.St <- scMergeSubClust(sc10x.St,i="St.dws.sc",g=c("Endo","SM","Fib","Leu"),nm="Merge") - -sc10x <- scMerge(sc10x,sc10x.Epi,sc10x.St,i.1="Merge",i.2="Merge",nm="Merge_Epi.dws.sc_St.dws.sc") - -sc10x <- scMerge(sc10x,sc10x,sc10x.Epi.NE,i.1="Merge_Epi.dws.sc_St.dws.sc",i.2="NE",nm="Merge_Epi.dws.sc_St.dws.sc_NE") - -sc10x <- SetAllIdent(object=sc10x,id="Merge_Epi.dws.sc_St.dws.sc") -sc10x@ident <- factor(sc10x@ident,levels=c("BE","LE","OE_SCGB","OE_KRT13","Fib","SM","Endo","Leu")) -postscript("./analysis/tSNE/FINAL/tSNE_FINAL.eps") -plot <- TSNEPlot(object=sc10x,pt.size=2.5,do.return=TRUE,vector.friendly=FALSE) -plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) -plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) -plot(plot) -dev.off() - -scTables(sc10x,i.1="samples",i.2="Merge_Epi.dws.sc_St.dws.sc") -scTables(sc10x,i.1="samples",i.2="Merge_Epi.dws.sc_St.dws.sc_NE") -scTables(sc10x,i.1="Merge_Epi.dws.sc_St.dws.sc_NE",i.2="Merge_Epi.dws.sc_St.dws.sc") - -save(list=ls(pattern="sc10x.Stress"),file="./analysis/sc10x.Stress.Rda") -rm(list=ls(pattern="sc10x.Stress")) -save(list=ls(pattern="^genes.deg"),file="./analysis/DEG.Rda") -rm(list=ls(pattern="^genes.deg")) -save(list=ls(pattern="sc10x.Epi"),file="./analysis/sc10x.Epi.Rda") -rm(list=ls(pattern="^sc10x.Epi")) -save(list=ls(pattern="sc10x.St"),file="./analysis/sc10x.St.Rda") -rm(list=ls(pattern="sc10x.St")) -save(list=ls(pattern="^sc10x"),file="./analysis/sc10x.Rda") -rm(list=ls(pattern="^sc10x")) -save.image(file="./analysis/Data.RData") diff --git a/r.scripts/sc-TissueMapper_RUN.D3.R b/r.scripts/sc-TissueMapper_RUN.D3.R deleted file mode 100644 index f06a90bc87ee94b20daad5fb7ccd8cfe89f9303b..0000000000000000000000000000000000000000 --- a/r.scripts/sc-TissueMapper_RUN.D3.R +++ /dev/null @@ -1,498 +0,0 @@ -gc() -library(methods) -library(optparse) -library(Seurat) -library(readr) -library(fBasics) -library(pastecs) -library(qusage) -library(RColorBrewer) - -source("../r.scripts/sc-TissueMapper.R") - -#Create folder structure -setwd("../") -if (!dir.exists("./analysis")){ - dir.create("./analysis") -} -if (!dir.exists("./analysis/qc")){ - dir.create("./analysis/qc") -} -if (!dir.exists("./analysis/qc/cc")){ - dir.create("./analysis/qc/cc") -} -if (!dir.exists("./analysis/tSNE")){ - dir.create("./analysis/tSNE") -} -if (!dir.exists("./analysis/tSNE/pre.stress")){ - dir.create("./analysis/tSNE/pre.stress") -} -if (!dir.exists("./analysis/pca")){ - dir.create("./analysis/pca") -} -if (!dir.exists("./analysis/pca/stress")){ - dir.create("./analysis/pca/stress") -} -if (!dir.exists("./analysis/violin")){ - dir.create("./analysis/violin") -} -if (!dir.exists("./analysis/violin/stress")){ - dir.create("./analysis/violin/stress") -} -if (!dir.exists("./analysis/table")){ - dir.create("./analysis/table") -} -if (!dir.exists("./analysis/tSNE/post.stress")){ - dir.create("./analysis/tSNE/post.stress") -} -if (!dir.exists("./analysis/cor")){ - dir.create("./analysis/cor") -} -if (!dir.exists("./analysis/tSNE/lin")){ - dir.create("./analysis/tSNE/lin") -} -if (!dir.exists("./analysis/tSNE/epi")){ - dir.create("./analysis/tSNE/epi") -} -if (!dir.exists("./analysis/tSNE/st")){ - dir.create("./analysis/tSNE/st") -} -if (!dir.exists("./analysis/tSNE/merge")){ - dir.create("./analysis/tSNE/merge") -} -if (!dir.exists("./analysis/pca/ne")){ - dir.create("./analysis/pca/ne") -} -if (!dir.exists("./analysis/tSNE/ne")){ - dir.create("./analysis/tSNE/ne") -} -if (!dir.exists("./analysis/violin/ne")){ - dir.create("./analysis/violin/ne") -} -if (!dir.exists("./analysis/tSNE/FINAL")){ - dir.create("./analysis/tSNE/FINAL") -} -if (!dir.exists("./analysis/deg")){ - dir.create("./analysis/deg") -} -if (!dir.exists("./analysis/cca")){ - dir.create("./analysis/cca") -} -if (!dir.exists("./analysis/diy")){ - dir.create("./analysis/diy") -} - -#Retrieve command-line options -option_list=list( - make_option("--p",action="store",default="DPrF",type='character',help="Project Name"), - make_option("--g",action="store",default="ALL",type='character',help="Group To analyze"), - make_option("--lg",action="store",default=500,type='integer',help="Threshold for cells with minimum genes"), - make_option("--hg",action="store",default=2500,type='integer',help="Threshold for cells with maximum genes"), - make_option("--lm",action="store",default=0,type='numeric',help="Threshold for cells with minimum %mito genes"), - make_option("--hm",action="store",default=0.1,type='numeric',help="Threshold for cells with maximum %mito genes"), - make_option("--lx",action="store",default=0.0125,type='numeric',help="x low threshold for hvg selection"), - make_option("--hx",action="store",default=3.5,type='numeric',help="x high threshold for hvg selection"), - make_option("--ly",action="store",default=0.5,type='numeric',help="y low threshold for hvg selection"), - make_option("--cc",action="store",default=TRUE,type='logical',help="Scale cell cycle?"), - make_option("--pc",action="store",default=25,type='integer',help="Number of PCs to cacluate"), - make_option("--hpc",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, pre-stress"), - make_option("--res.prestress",action="store",default=1,type='numeric',help="Resolution to cluster, pre-stress"), - make_option("--st",action="store",default="TRUE",type='logical',help="Remove stressed cells?"), - make_option("--stg",action="store",default="dws",type='character',help="Geneset to use for stress ID"), - #make_option("--hpc.poststress",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, post-stress"), - make_option("--res.poststress",action="store",default=0.2,type='numeric',help="Resolution to cluster, post-stress"), - make_option("--ds",action="store",default=10000,type='integer',help="Number of cells to downsample"), - #make_option("--hpc.epi",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, Epi"), - make_option("--res.epi",action="store",default=0.2,type='numeric',help="Resolution to cluster, Epi"), - #make_option("--hpc.st",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, St"), - make_option("--res.st",action="store",default=0.2,type='numeric',help="Resolution to cluster, St") - ) -opt=parse_args(OptionParser(option_list=option_list)) -rm(option_list) -if (opt$lm==0){opt$lm=-Inf} - -sc10x.D17 <- scLoad("D17PrF",sub=TRUE) -sc10x.D27 <- scLoad("D27PrF",sub=TRUE) -sc10x.D35 <- scLoad("D35PrF",sub=TRUE) - -if (opt$cc==TRUE){ - results <- scCellCycle(sc10x.D17) - sc10x.D17 <- results[[1]] - genes.s <- results[[2]] - genes.g2m <- results[[3]] - rm(results) - results <- scCellCycle(sc10x.D27) - sc10x.D27 <- results[[1]] - genes.s <- results[[2]] - genes.g2m <- results[[3]] - rm(results) - results <- scCellCycle(sc10x.D35) - sc10x.D35 <- results[[1]] - genes.s <- results[[2]] - genes.g2m <- results[[3]] - rm(results) -} else { - genes.s="" - genes.g2m="" -} - -results <- scQC(sc10x.D17,lg=opt$lg,hg=opt$hg,lm=opt$lm,hm=opt$hm) -sc10x.D17 <- results[[1]] -counts.cell.raw.D17 <- results[[2]] -counts.gene.raw.D17 <- results[[3]] -counts.cell.filtered.D17 <- results[[4]] -counts.gene.filtered.D17 <- results[[5]] -rm(results) - -results <- scQC(sc10x.D27,lg=opt$lg,hg=opt$hg,lm=opt$lm,hm=opt$hm) -sc10x.D27 <- results[[1]] -counts.cell.raw.D27 <- results[[2]] -counts.gene.raw.D27 <- results[[3]] -counts.cell.filtered.D27 <- results[[4]] -counts.gene.filtered.D27 <- results[[5]] -rm(results) - -results <- scQC(sc10x.D35,lg=opt$lg,hg=opt$hg,lm=opt$lm,hm=opt$hm) -sc10x.D35 <- results[[1]] -counts.cell.raw.D35 <- results[[2]] -counts.gene.raw.D35 <- results[[3]] -counts.cell.filtered.D35 <- results[[4]] -counts.gene.filtered.D35 <- results[[5]] -rm(results) - -results <- sc3CCA(sc10x.D17,sc10x.D27,sc10x.D35,nm.1="D17",nm.2="D27",nm.3="D35",cc=opt$cc) -sc10x <- results[[1]] -genes.hvg.cca <- results[[2]] -rm(results) - -rm(sc10x.D17) -rm(sc10x.D27) -rm(sc10x.D35) -save.image(file="./analysis/Aligned.RData") - -#results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="pre.stress",cca=TRUE) -#sc10x <- results[[1]] -#genes.hvg <- results[[2]] -#pc.use <- results[[3]] -#rm(results) - -pc.use <- 15 -sc10x <- scCluster(sc10x,pc.use=pc.use,res.use=opt$res.prestress,folder="pre.stress",red="cca.aligned") - -if (opt$st==TRUE){ - results <- scStress(sc10x,stg=opt$stg,res.use=opt$res.prestress,pc.use=pc.use,cut=0.95) - sc10x <- results[[1]] - counts.cell.filtered.stress <- results[[2]] - sc10x.Stress <- results[[3]] - rm(results) - - #results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="post.stress") - #sc10x <- results[[1]] - #genes.hvg.poststress <- results[[2]] - #pc.use.poststress <- results[[3]] - #rm(results) - - sc10x <- scCluster(sc10x,pc.use=pc.use,res.use=opt$res.poststress,folder="post.stress",red="cca.aligned") -} - -gene.set1 <- read_delim("./genesets/genes.deg.St.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "St" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.Epi.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Epi" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -gc() -results <- scQuSAGE(sc10x,gs=gene.set,res.use=opt$res.poststress,ds=opt$ds,nm="Lin",folder="lin") -sc10x <- results[[1]] -results.cor.Lin <- results[[2]] -results.clust.Lin.id <- results[[3]] -rm(results) -rm(gene.set) - -sc10x.Epi <- scSubset(sc10x,i="Lin",g="Epi") -if (any(levels(sc10x@ident)=="Unknown")){ - sc10x.St <- scSubset(sc10x,i="Lin",g=c("St","Unknown")) -} else { - sc10x.St <- scSubset(sc10x,i="Lin",g="St") -} - -sc10x.Epi <- scCluster(sc10x.Epi,pc.use=pc.use,res.use=opt$res.epi,folder="epi",red="cca.aligned") - -gene.set1 <- read_delim("./genesets/genes.deg.BE.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "BE" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.LE.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "LE" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.OE1.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "OE_SCGB" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.OE2.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "OE_KRT13" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -gc() -results <- scQuSAGE(sc10x.Epi,gs=gene.set,res.use=opt$res.epi,ds=opt$ds,nm="Epi.dws.sc",folder="epi") -sc10x.Epi <- results[[1]] -results.cor.Epi.dws.sc <- results[[2]] -results.clust.Epi.dws.sc.id <- results[[3]] -rm(results) -rm(gene.set) - -gene.set1 <- read_csv("./genesets/Basal cells-signature-genes.csv") -gene.set1 <- gene.set1[,2] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "BC" -gene.set <- c(gene.set1) -gene.set1 <- read_csv("./genesets/Normal AT2 cells-signature-genes.csv") -gene.set1 <- gene.set1[,2] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "AT2" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_csv("./genesets/Club_Goblet cells-signature-genes.csv") -gene.set1 <- gene.set1[,2] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "CGC" -gene.set<- c(gene.set,gene.set1) -rm(gene.set1) -gc() -results.cor.Epi.lgea <- scQuSAGEsm(sc10x.Epi,gs=gene.set,ds=opt$ds,nm="Epi.dws.sc",folder="lgea") -rm(gene.set) - -sc10x.St <- scCluster(sc10x.St,pc.use=pc.use,res.use=opt$res.epi,folder="st",red="cca.aligned") - -gene.set1 <- read_delim("./genesets/genes.deg.Fib.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Fib" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.SM.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "SM" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.Endo.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Endo" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/genes.deg.Leu.csv",",",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- gene.set1[1] -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "Leu" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -gc() -results <- scQuSAGE(sc10x.St,gs=gene.set,res.use=opt$res.st,ds=opt$ds,nm="St.dws.sc",folder="st") -sc10x.St <- results[[1]] -results.cor.St.dws.sc <- results[[2]] -results.clust.St.dws.sc.id <- results[[3]] -rm(results) -rm(gene.set) - -sc10x.Epi.NE <- scNE(sc10x.Epi,neg="dws"cut=0.95) - -sc10x.Epi <- scMergeSubClust(sc10x.Epi,i="Epi.dws.sc",g=c("BE","LE","OE_SCGB","OE_KRT13"),nm="Merge") - -sc10x.St <- scMergeSubClust(sc10x.St,i="St.dws.sc",g=c("Endo","SM","Fib","Leu"),nm="Merge") - -sc10x <- scMerge(sc10x,sc10x.Epi,sc10x.St,i.1="Merge",i.2="Merge",nm="Merge_Epi.dws.sc_St.dws.sc") - -sc10x <- scMerge(sc10x,sc10x,sc10x.Epi.NE,i.1="Merge_Epi.dws.sc_St.dws.sc",i.2="NE",nm="Merge_Epi.dws.sc_St.dws.sc_NE") - -sc10x <- SetAllIdent(object=sc10x,id="Merge_Epi.dws.sc_St.dws.sc") -sc10x@ident <- factor(sc10x@ident,levels=c("BE","LE","OE_SCGB","OE_KRT13","Fib","SM","Endo","Leu")) -postscript("./analysis/tSNE/FINAL/tSNE_FINAL.eps") -plot <- TSNEPlot(object=sc10x,pt.size=2.5,do.return=TRUE,vector.friendly=FALSE) -plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) -plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) -plot(plot) -dev.off() - -scTables(sc10x,i.1="samples",i.2="Merge_Epi.dws.sc_St.dws.sc") -scTables(sc10x,i.1="samples",i.2="Merge_Epi.dws.sc_St.dws.sc_NE") -scTables(sc10x,i.1="Merge_Epi.dws.sc_St.dws.sc_NE",i.2="Merge_Epi.dws.sc_St.dws.sc") - -genes.deg.Stress <- scDEG(sc10x.Stress,i="Stress",g.1="Stress",g.2="ALL",pct=0.5,t=5) - -genes.deg.Epi <- scDEG(sc10x,i="Lin",g.1="Epi",g.2="St",t=2) -genes.deg.St <- scDEG(sc10x,i="Lin",g.1="St",g.2="Epi",t=2) - -genes.deg.BE <- scDEG(sc10x.Epi.NE,i="Epi.dws.sc",g.1="BE",g.2=c("LE","OE_SCGB","OE_KRT13"),pct=0.25,t=2) -genes.deg.LE <- scDEG(sc10x.Epi.NE,i="Epi.dws.sc",g.1="LE",g.2=c("BE","LE","OE_SCGB"),pct=0.25,t=2) -genes.deg.OE_SCGB <- scDEG(sc10x.Epi.NE,i="Epi.dws.sc",g.1="OE_SCGB",g.2=c("BE","LE","OE_KRT13"),pct=0.25,t=2) -genes.deg.OE_KRT13 <- scDEG(sc10x.Epi.NE,i="Epi.dws.sc",g.1="OE_KRT13",g.2=c("BE","LE","OE_SCGB"),pct=0.25,t=2) - -genes.deg.NE <- scDEG(sc10x.Epi.NE,i="NE",g.1="NE",g.2="ALL",pct=0.25,t=2) - -genes.deg.Fib <- scDEG(sc10x.St,i="St.dws.sc",g.1="Fib",g.2=c("SM","Endo","Leu"),pct=0.25,t=2) -genes.deg.SM <- scDEG(sc10x.St,i="St.dws.sc",g.1="SM",g.2=c("Fib","Endo","Leu"),pct=0.25,t=2) -genes.deg.Endo <- scDEG(sc10x.St,i="St.dws.sc",g.1="Endo",g.2=c("Fib","SM","Leu"),pct=0.25,t=2) -genes.deg.Leu <- scDEG(sc10x.St,i="St.dws.sc",g.1="Leu",g.2=c("Fib","SM","Endo"),pct=0.25,t=2) - -genes.deg.BE.unique <- setdiff(rownames(genes.deg.BE),Reduce(union,list(rownames(genes.deg.St),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.LE.unique <- setdiff(rownames(genes.deg.LE),Reduce(union,list(rownames(genes.deg.St),rownames(genes.deg.BE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.OE_SCGB.unique <- setdiff(rownames(genes.deg.OE_SCGB),Reduce(union,list(rownames(genes.deg.St),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.OE_KRT13.unique <- setdiff(rownames(genes.deg.OE_KRT13),Reduce(union,list(rownames(genes.deg.St),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.NE.unique <- setdiff(rownames(genes.deg.NE),Reduce(union,list(rownames(genes.deg.St),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.Fib.unique <- setdiff(rownames(genes.deg.Fib),Reduce(union,list(rownames(genes.deg.Epi),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.SM),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.SM.unique <- setdiff(rownames(genes.deg.SM),Reduce(union,list(rownames(genes.deg.Epi),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.Endo),rownames(genes.deg.Leu)))) -genes.deg.Endo.unique <- setdiff(rownames(genes.deg.Endo),Reduce(union,list(rownames(genes.deg.Epi),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Leu)))) -genes.deg.Leu.unique <- setdiff(rownames(genes.deg.Leu),Reduce(union,list(rownames(genes.deg.Epi),rownames(genes.deg.BE),rownames(genes.deg.LE),rownames(genes.deg.OE_SCGB),rownames(genes.deg.OE_KRT13),rownames(genes.deg.NE),rownames(genes.deg.Fib),rownames(genes.deg.SM),rownames(genes.deg.Endo)))) - -genes.deg.5 <- c(genes.deg.BE.unique[1:5],genes.deg.LE.unique[1:5],genes.deg.OE_SCGB.unique[1:5],genes.deg.OE_KRT13.unique[1:5],genes.deg.NE.unique[1:5],genes.deg.Fib.unique[1:5],genes.deg.SM.unique[1:5],genes.deg.Endo.unique[1:5],genes.deg.Leu.unique[1:5]) -genes.deg.5 <- rev(genes.deg.5) -genes.deg.10 <- c(genes.deg.BE.unique[1:10],genes.deg.LE.unique[1:10],genes.deg.OE_SCGB.unique[1:10],genes.deg.OE_KRT13.unique[1:10],genes.deg.NE.unique[1:10],genes.deg.Fib.unique[1:10],genes.deg.SM.unique[1:10],genes.deg.Endo.unique[1:10],genes.deg.Leu.unique[1:10]) -genes.deg.10 <- rev(genes.deg.10) - - -sc10x <- SetAllIdent(object=sc10x,id="Merge_Epi.dws.sc_St.dws.sc_NE") -sc10x <- SetAllIdent(object=sc10x,id="Merge_Epi.dws.sc_St.dws.sc_NE") -sc10x@ident <- factor(sc10x@ident,levels=c("BE","LE","OE_SCGB","OE_KRT13","NE","Fib","SM","Endo","Leu")) -postscript("./analysis/deg/Dot.DEG.eps",paper="special",width=10,height=5,horizontal=FALSE) -DotPlot(sc10x,genes.deg.5,x.lab.rot=TRUE,plot.legend=TRUE,dot.scale=4) -dev.off() -postscript("./analysis/deg/Heatmap.DEG.eps",paper="special",width=10,height=5,horizontal=FALSE) -plot(DoHeatmap(sc10x,genes.use=genes.deg.10,slim.col.label=TRUE,group.label.rot=TRUE,group.spacing=0.25,cex.row=2.5)) -dev.off() - -for (i in ls(pattern="^genes.deg*unique")){ - postscript(paste0("./analysis/deg/Violin.",i,".eps"),paper="special",width=10,height=5,horizontal=FALSE) - plot(VlnPlot(sc10x,features.plot=get(i)[1:10],nCol=5,point.size.use=0.1,size.title.use=15,x.lab.rot=TRUE)) - dev.off() - postscript(paste0("./analysis/deg/Ridge.",i,".eps"),paper="special",width=10,height=5,horizontal=FALSE) - plot(RidgePlot(sc10x,features.plot=get(i)[1:10],nCol=5,size.x.use=10,size.title.use=15)) - dev.off() - postscript(paste0("./analysis/deg/Heatmap.",i,".eps"),paper="special",width=10,height=5,horizontal=FALSE) - plot(DoHeatmap(sc10x,genes.use=get(i),slim.col.label=TRUE,group.label.rot=TRUE,group.spacing=0.25,cex.row=2.5)) - dev.off() -} - -sc10x.Stress <- SetAllIdent(object=sc10x.Stress,id="Stress") -postscript("./analysis/deg/Violin.Stress.eps",paper="special",width=10,height=5,horizontal=FALSE) -plot(VlnPlot(sc10x.Stress,features.plot=rownames(genes.deg.Stress)[1:10],nCol=5,point.size.use=0.1,size.title.use=15,x.lab.rot=TRUE)) -dev.off() -postscript("./analysis/deg/Ridge.Stress.eps",paper="special",width=10,height=5,horizontal=FALSE) -plot(RidgePlot(sc10x.Stress,features.plot=rownames(genes.deg.Stress)[1:10],nCol=5,size.x.use=10,size.title.use=15)) -dev.off() -postscript("./analysis/deg/Heatmap.Stress.eps",paper="special",width=10,height=5,horizontal=FALSE) -plot(DoHeatmap(sc10x.Stress,genes.use=rownames(genes.deg.Stress),slim.col.label=TRUE,group.label.rot=TRUE,group.spacing=0.25,cex.row=2.5)) -dev.off() - -sc10x.Epi.NE <- SetAllIdent(object=sc10x.Epi.NE,id="NE") -postscript("./analysis/deg/Violin.NE.eps",paper="special",width=10,height=5,horizontal=FALSE) -plot(VlnPlot(sc10x.Epi.NE,features.plot=rownames(genes.deg.NE)[1:10],nCol=5,point.size.use=0.1,size.title.use=15,x.lab.rot=TRUE)) -dev.off() -postscript("./analysis/deg/Ridge.NE.eps",paper="special",width=10,height=5,horizontal=FALSE) -plot(RidgePlot(sc10x.Epi.NE,features.plot=rownames(genes.deg.NE)[1:10],nCol=5,size.x.use=10,size.title.use=15)) -dev.off() - -for (i in ls(pattern="^genes.deg")){ - write.table(get(i),file=paste0("./analysis/deg/",i,".csv"),row.names=TRUE,col.names=NA,append=FALSE,sep=",") -} - -sctSNECustCol(sc10x,i="Lin",bl="Epi",rd="St",file="Aggr") -sctSNECustCol(sc10x,i="Merge_Epi.dws.sc_St.dws.sc",bl=c("BE","LE","OE_SCGB","OE_KRT13"),rd=c("Fib","SM","Endo","Leu"),file="Aggr") -sctSNECustCol(sc10x.Epi,i="Epi.dws.sc",bl=c("BE","LE","OE_SCGB","OE_KRT13"),rd="",file="Aggr") -sctSNECustCol(sc10x.St,i="St.dws.sc",bl="",rd=c("Fib","SM","Endo","Leu"),file="Aggr") - -sctSNEbwCol(sc10x,i="res0.2",file="ALL",files="Aggr") -sctSNEbwCol(sc10x.Epi,i="res0.2",file="Epi",files="Aggr") -sctSNEbwCol(sc10x.St,i="res0.2",file="St",files="Aggr") -sctSNEbwCol(sc10x,i="Merge_Epi.dws.sc_St.dws.sc",file="ALL",files="Aggr") -sctSNEbwCol(sc10x.Epi,i="Epi.dws.sc",file="Epi",files="Aggr") -sctSNEbwCol(sc10x.St,i="St.dws.sc",file="St",files="Aggr") - -for (g in c("Epi","St")){ - sctSNEHighlight(sc10x,i="Lin",g=g,file="Aggr") -} -for (g in c("BE","LE","OE_SCGB","OE_KRT13")){ - sctSNEHighlight(sc10x,i="Merge_Epi.dws.sc_St.dws.sc",g=g,file="Aggr") - sctSNEHighlight(sc10x.Epi,i="Epi.dws.sc",g=g,file="Aggr") -} -sctSNEHighlight(sc10x.Epi.NE,i="NE",g="NE",file="Aggr") -for (g in c("Fib","SM","Endo","Leu")){ - sctSNEHighlight(sc10x,i="Merge_Epi.dws.sc_St.dws.sc",g=g,file="Aggr") - sctSNEHighlight(sc10x.St,i="St.dws.sc",g=g,file="Aggr") -} -for (g in c("D17PrPzF_BE","D17PrPzF_LE","D17PrPzF_OE","D17PrPzF_FMSt","D17PrPzF_Via","D17PrTzF_Via","D27PrTzF_BE","D27PrTzF_LE","D27PrTzF_OE","D27PrTzF_Edn","D27PrTzF_StPDPNp","D27PrTzF_StPDPNn","D27PrPzF_Via","D27PrTzF_Via")){ - sctSNEHighlight(sc10x,i="samples",g=g,file="Aggr") -} -rm(g) - -scCustHeatmap(sc10x.Epi,i="Epi.dws.sub",gs=c(genes.deg.BE.unique,genes.deg.LE.unique,genes.deg.OE_SCGB.unique,genes.deg.OE_KRT13.unique),g=c("BE","LE","OE_SCGB","OE_KRT13")) -scCustHeatmap(sc10x.St,i="St.go",gs=c(genes.deg.Fib.unique,genes.deg.SM.unique,genes.deg.Endo.unique,genes.deg.Leu.unique),g=c("Fib","SM","Endo","Leu")) - - -sc10x.D27 <- scSubset(sc10x,i="samples",g=c("D27PrTzF_BE","D27PrTzF_LE","D27PrTzF_OE","D27PrTzF_Edn","D27PrTzF_StPDPNp","D27PrTzF_StPDPNn","D27PrPzF_Via","D27PrTzF_Via")) -sc10x.D27.Epi <- scSubset(sc10x.Epi,i="samples",g=c("D27PrTzF_BE","D27PrTzF_LE","D27PrTzF_OE","D27PrTzF_Edn","D27PrTzF_StPDPNp","D27PrTzF_StPDPNn","D27PrPzF_Via","D27PrTzF_Via")) -sc10x.D27.Epi.NE <- scSubset(sc10x.Epi.NE,i="samples",g=c("D27PrTzF_BE","D27PrTzF_LE","D27PrTzF_OE","D27PrTzF_Edn","D27PrTzF_StPDPNp","D27PrTzF_StPDPNn","D27PrPzF_Via","D27PrTzF_Via")) -sc10x.D27.St <- scSubset(sc10x.St,i="samples",g=c("D27PrTzF_BE","D27PrTzF_LE","D27PrTzF_OE","D27PrTzF_Edn","D27PrTzF_StPDPNp","D27PrTzF_StPDPNn","D27PrPzF_Via","D27PrTzF_Via")) - -sc10x.D27 <- RunTSNE(object=sc10x.D27,dims.use=1:pc.use.poststress,do.fast=TRUE) -sc10x.D27.Epi <- RunTSNE(object=sc10x.D27.Epi,dims.use=1:pc.use.poststress,do.fast=TRUE) -sc10x.D27.Epi.NE <- RunTSNE(object=sc10x.D27.Epi.NE,dims.use=1:pc.use.poststress,do.fast=TRUE) -sc10x.D27.St <- RunTSNE(object=sc10x.D27.St,dims.use=1:pc.use.poststress,do.fast=TRUE) - -sctSNECustCol(sc10x.D27,i="Lin",bl="Epi",rd="St",file="D27") -sctSNECustCol(sc10x.D27,i="Merge_Epi.dws.sc_St.dws.sc",bl=c("BE","LE","OE_SCGB","OE_KRT13"),rd=c("Fib","SM","Endo","Leu"),file="D27") -sctSNECustCol(sc10x.D27.Epi,i="Epi.dws.sc",bl=c("BE","LE","OE_SCGB","OE_KRT13"),rd="",file="D27.Epi") -sctSNECustCol(sc10x.D27.St,i="St.dws.sc",bl="",rd=c("Fib","SM","Endo","Leu"),file="D27.St") - -sctSNEbwCol(sc10x.D27,i="res0.2",file="ALL",files="D27") -sctSNEbwCol(sc10x.D27.Epi,i="res0.2",file="Epi",files="D27") -sctSNEbwCol(sc10x.D27.St,i="res0.2",file="St",files="D27") -sctSNEbwCol(sc10x.D27,i="Merge_Epi.dws.sc_St.dws.sc",file="ALL",files="D27") -sctSNEbwCol(sc10x.D27.Epi,i="Epi.dws.sc",file="Epi",files="D27") -sctSNEbwCol(sc10x.D27.St,i="St.dws.sc",file="St",files="D27") - -for (g in c("Epi","St")){ - sctSNEHighlight(sc10x.D27,i="Lin",g=g,file="D27") -} -for (g in c("BE","LE","OE_SCGB","OE_KRT13")){ - sctSNEHighlight(sc10x.D27,i="Merge_Epi.dws.sc_St.dws.sc",g=g,file="D27") - sctSNEHighlight(sc10x.D27.Epi,i="Epi.dws.sc",g=g,file="D27") -} -sctSNEHighlight(sc10x.D27.Epi.NE,i="NE",g="NE",file="D27") -for (g in c("Fib","SM","Endo","Leu")){ - sctSNEHighlight(sc10x.D27,i="Merge_Epi.dws.sc_St.dws.sc",g=g,file="D27") - sctSNEHighlight(sc10x.D27.St,i="St.dws.sc",g=g,file="D27") -} -for (g in c("D27PrTzF_BE","D27PrTzF_LE","D27PrTzF_OE","D27PrTzF_Edn","D27PrTzF_StPDPNp","D27PrTzF_StPDPNn","D27PrPzF_Via","D27PrTzF_Via")){ - sctSNEHighlight(sc10x.D27,i="samples",g=g,file="D27") -} -rm(g) - -sc10x.D27 <- SetAllIdent(object=sc10x.D27,id="samples") -cell <- names(sc10x.D27@ident[grep("^D27PrTzF_StPDPN",as.character(sc10x.D27@ident))]) -sc10x.D27 <- SetIdent(object=sc10x.D27,cells.use=cell,ident.use="D27PrTzF_FMSt") -rm(cell) -sctSNEHighlight(sc10x.D27,i="current",g="D27PrTzF_FMSt",file="D27") - - -save(list=ls(pattern="^sc10x.D27"),file="./analysis/sc10x.D27.Rda") -rm(list=ls(pattern="^sc10x.D27")) -save(list=ls(pattern="sc10x.Stress"),file="./analysis/sc10x.Stress.Rda") -rm(list=ls(pattern="sc10x.Stress")) -save(list=ls(pattern="^genes.deg"),file="./analysis/DEG.Rda") -rm(list=ls(pattern="^genes.deg")) -save(list=ls(pattern="sc10x.Epi"),file="./analysis/sc10x.Epi.Rda") -rm(list=ls(pattern="^sc10x.Epi")) -save(list=ls(pattern="sc10x.St"),file="./analysis/sc10x.St.Rda") -rm(list=ls(pattern="sc10x.St")) -save(list=ls(pattern="^sc10x"),file="./analysis/sc10x.Rda") -rm(list=ls(pattern="^sc10x")) -save.image(file="./analysis/Data.RData")