diff --git a/bash.scripts/sc_TissueMapper-PdPbPc.sh b/bash.scripts/sc_TissueMapper-PdPbPc.sh index 7422a5c2e571b2a565245d3b48b766f68672690c..3cbf55f5bf89c96ea43c666b202d27bb2dd61e9e 100644 --- a/bash.scripts/sc_TissueMapper-PdPbPc.sh +++ b/bash.scripts/sc_TissueMapper-PdPbPc.sh @@ -8,7 +8,11 @@ #SBATCH --mail-type ALL #SBATCH --mail-user gervaise.henry@utsouthwestern.edu -module load R/3.4.1-gccmkl +sh sc_LinkData.sh Pd Pd +sh sc_LinkData.sh Pb Pb +sh sc_LinkData.sh Pc Pc + +module load R/3.4.1-gccmkl_20181025 Rscript ../r.scripts/sc-TissueMapper_RUN.PdPbPc.Pd.R & Rscript ../r.scripts/sc-TissueMapper_RUN.PdPbPc.Pb.R & diff --git a/r.scripts/sc-TissueMapper_RUN.PdPbPc.R b/r.scripts/sc-TissueMapper_RUN.PdPbPc.R index 24eeb91a0a54e00bd2cfcc4e8b5edf1df1cd2288..aeb3cfbbfa218f5154801c3387c886828eff3acb 100755 --- a/r.scripts/sc-TissueMapper_RUN.PdPbPc.R +++ b/r.scripts/sc-TissueMapper_RUN.PdPbPc.R @@ -58,12 +58,18 @@ if (!dir.exists("./analysis/tSNE/lin")){ if (!dir.exists("./analysis/tSNE/epi")){ dir.create("./analysis/tSNE/epi") } +if (!dir.exists("./analysis/tSNE/n-epi")){ + dir.create("./analysis/tSNE/n-epi") +} if (!dir.exists("./analysis/tSNE/st")){ dir.create("./analysis/tSNE/st") } if (!dir.exists("./analysis/tSNE/leu")){ dir.create("./analysis/tSNE/leu") } +if (!dir.exists("./analysis/tSNE/t")){ + dir.create("./analysis/tSNE/t") +} if (!dir.exists("./analysis/tSNE/merge")){ dir.create("./analysis/tSNE/merge") } @@ -126,21 +132,27 @@ for (i in c("Pb","Pc","Pd")){ if (!dir.exists(paste0("./analysis/tSNE/post.stress/",i))){ dir.create(paste0("./analysis/tSNE/post.stress/",i)) } - if (!dir.exists(paste0("./analysis/tSNE/lin/",i))){ + if (!dir.exists(paste0("./analysis/tSNE/lin/",i))){ dir.create(paste0("./analysis/tSNE/lin/",i)) } if (!dir.exists(paste0("./analysis/tSNE/epi/",i))){ dir.create(paste0("./analysis/tSNE/epi/",i)) } + if (!dir.exists(paste0("./analysis/tSNE/n-epi/",i))){ + dir.create(paste0("./analysis/tSNE/n-epi/",i)) + } if (!dir.exists(paste0("./analysis/tSNE/st/",i))){ dir.create(paste0("./analysis/tSNE/st/",i)) } if (!dir.exists(paste0("./analysis/tSNE/leu/",i))){ dir.create(paste0("./analysis/tSNE/leu/",i)) } - + if (!dir.exists(paste0("./analysis/tSNE/t/",i))){ + dir.create(paste0("./analysis/tSNE/t/",i)) + } + sc10x <- get(paste0("sc10x.",i)) - + sc10x <- scCluster(sc10x,pc.use=opt$acca,res.use=opt$res.poststress,folder=paste0("post.stress/",i),red="cca.aligned") gene.set1 <- read_delim("./genesets/DEG_Epi_5FC.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=FALSE) @@ -153,8 +165,8 @@ for (i in c("Pb","Pc","Pd")){ gene.set <- c(gene.set,gene.set1) rm(gene.set1) gc() - min.all <- min(table(sc10x@meta.data[,paste0("res",0.5)])) - results <- scQuSAGE(sc10x,gs=gene.set,res.use=opt$res.poststress,ds=0,nm="Lin",folder=paste0("lin/",i)) + min.all <- min(table(sc10x@meta.data[,paste0("res",opt$res.poststress)])) + results <- scQuSAGE(sc10x,gs=gene.set,res.use=opt$res.poststress,ds=min.all,nm="Lin",folder=paste0("lin/",i)) sc10x <- results[[1]] results.cor.Lin <- results[[2]] results.clust.Lin.id <- results[[3]] @@ -164,15 +176,12 @@ for (i in c("Pb","Pc","Pd")){ sc10x <- SetAllIdent(object=sc10x,id="Lin") sc10x.Epi <- scSubset(sc10x,i="Lin",g="Epi") if (any(levels(sc10x@ident)=="Unknown")){ - sc10x.St <- scSubset(sc10x,i="Lin",g=c("St","Unknown")) + sc10x.nEpi <- scSubset(sc10x,i="Lin",g=c("St","Unknown")) } else { - sc10x.St <- scSubset(sc10x,i="Lin",g="St") + sc10x.nEpi <- scSubset(sc10x,i="Lin",g="St") } sc10x.Epi <- scCluster(sc10x.Epi,pc.use=opt$acca,res.use=opt$res.poststress,folder=paste0("epi/",i),red="cca.aligned") - - #sc10x.Epi <- SetAllIdent(object=sc10x.Epi,id=paste0("res",opt$res.poststress)) - #sc10x.Epi <- BuildClusterTree(sc10x.Epi,do.reorder=TRUE,reorder.numeric=TRUE,do.plot=FALSE) sc10x.Epi <- StashIdent(object=sc10x.Epi,save.name=paste0("res",opt$res.poststress)) sc10x.Epi <- RunTSNE(object=sc10x.Epi,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) @@ -213,29 +222,25 @@ for (i in c("Pb","Pc","Pd")){ rm(gene.set1) gc() min.epi <- min(table(sc10x.Epi@meta.data[,paste0("res",opt$res.poststress)])) - results <- scQuSAGE(sc10x.Epi,gs=gene.set,res.use=opt$res.poststress,ds=0,nm="Epi.dws.sc",folder=paste0("epi/",i)) + results <- scQuSAGE(sc10x.Epi,gs=gene.set,res.use=opt$res.poststress,ds=min.epi,nm="Epi.dws.sc",folder=paste0("epi/",i)) sc10x.Epi <- results[[1]] results.cor.Epi <- results[[2]] results.clust.Epi.id <- results[[3]] rm(results) rm(gene.set) + sc10x.nEpi <- scCluster(sc10x.nEpi,pc.use=opt$acca,res.use=opt$res.poststress,folder=paste0("n-epi/",i),red="cca.aligned") + sc10x.nEpi <- StashIdent(object=sc10x.nEpi,save.name=paste0("res",opt$res.poststress)) - sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=opt$res.poststress,folder=paste0("st/",i),red="cca.aligned") - - #sc10x.St <- SetAllIdent(object=sc10x.St,id=paste0("res",opt$res.poststress)) - #sc10x.St <- BuildClusterTree(sc10x.St,do.reorder=TRUE,reorder.numeric=TRUE,do.plot=FALSE) - sc10x.St <- StashIdent(object=sc10x.St,save.name=paste0("res",opt$res.poststress)) - - sc10x.St <- RunTSNE(object=sc10x.St,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) - postscript(paste0("./analysis/tSNE/st/",i,"/tSNE_Sample.eps")) - plot <- TSNEPlot(object=sc10x.St,group.by="samples",pt.size=2.5,do.return=TRUE,vector.friendly=FALSE) + sc10x.nEpi <- RunTSNE(object=sc10x.nEpi,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) + postscript(paste0("./analysis/tSNE/n-epi/",i,"/tSNE_Sample.eps")) + plot <- TSNEPlot(object=sc10x.nEpi,group.by="samples",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() - postscript(paste0("./analysis/tSNE/st/",i,"/tSNE_res",opt$res.poststress,".eps")) - plot <- TSNEPlot(object=sc10x.St,pt.size=5,do.label=TRUE,label.size=10,do.return=TRUE,vector.friendly=FALSE) + postscript(paste0("./analysis/tSNE/n-epi/",i,"/tSNE_res",opt$res.poststress,".eps")) + plot <- TSNEPlot(object=sc10x.nEpi,pt.size=5,do.label=TRUE,label.size=10,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) @@ -257,29 +262,39 @@ for (i in c("Pb","Pc","Pd")){ gene.set1 <- as.list(gene.set1) names(gene.set1) <- "Fib" gene.set <- c(gene.set,gene.set1) - - genes.leu <- read_excel("genesets/40425_2017_215_MOESM1_ESM.xlsx",sheet="S3. Candidate markers") - leu <- as.list(unique(genes.leu[,2]))$Cell - leu <- leu[-c(1,3,4,7:9,14,15,17:18,20:21,23:30)] - genes.leu <- genes.leu[genes.leu$Selected==1,] - genes.leu <- genes.leu[unlist(genes.leu[,2]) %in% unlist(leu),] - gene.set1 <- split(genes.leu[,1], genes.leu[,2]) - gene.set1 <- lapply(gene.set1,unname) - gene.set1 <- lapply(gene.set1,unlist) - gene.set <- c(gene.set,gene.set1) - rm(gene.set1) gc() - min.st <- min(table(sc10x.St@meta.data[,paste0("res",opt$res.poststress)])) - results <- scQuSAGE(sc10x.St,gs=gene.set,res.use=opt$res.poststress,ds=0,nm="St.dws.sc",folder=paste0("st/",i)) - sc10x.St <- results[[1]] + min.nepi <- min(table(sc10x.nEpi@meta.data[,paste0("res",opt$res.poststress)])) + results <- scQuSAGE(sc10x.nEpi,gs=gene.set,res.use=opt$res.poststress,ds=min.nepi,nm="St.dws.sc",folder=paste0("n-epi/",i)) + sc10x.nEpi <- results[[1]] results.cor.St <- results[[2]] results.clust.St.id <- results[[3]] rm(results) rm(gene.set) - sc10x.St <- SetAllIdent(object=sc10x.St,id="St.dws.sc") - sc10x.Leu <- scSubset(sc10x.St,i="St.dws.sc",g=leu[leu %in% unique(sc10x.St@ident)]) + sc10x.nEpi <- SetAllIdent(object=sc10x.nEpi,id="St.dws.sc") + sc10x.St <- scSubset(sc10x.nEpi,i="St.dws.sc",g=c("Fib","SM","Endo")[c("Fib","SM","Endo") %in% unique(sc10x.nEpi@ident)]) + sc10x.Leu <- scSubset(sc10x.nEpi,i="St.dws.sc",g="Unknown") + + sc10x.St <- RunTSNE(object=sc10x.St,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) + postscript(paste0("./analysis/tSNE/st/",i,"/tSNE_Sample.eps")) + plot <- TSNEPlot(object=sc10x.St,group.by="samples",pt.size=2.5,do.return=TRUE,vector.friendly=FALSE) + plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) + plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) + plot(plot) + dev.off() + postscript(paste0("./analysis/tSNE/st/",i,"/tSNE_St.dws.sc.eps")) + plot <- TSNEPlot(object=sc10x.St,pt.size=5,do.label=TRUE,label.size=10,do.return=TRUE,vector.friendly=FALSE) + plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) + plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) + plot(plot) + dev.off() + rm(plot) + + opt$res.leu <- 2 + + sc10x.Leu <- scCluster(sc10x.Leu,pc.use=opt$acca,res.use=opt$res.leu,folder=paste0("leu/",i),red="cca.aligned") + sc10x.Leu <- StashIdent(object=sc10x.Leu,save.name=paste0("res",opt$res.leu)) sc10x.Leu <- RunTSNE(object=sc10x.Leu,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) postscript(paste0("./analysis/tSNE/leu/",i,"/tSNE_Sample.eps")) @@ -288,7 +303,7 @@ for (i in c("Pb","Pc","Pd")){ plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) plot(plot) dev.off() - postscript(paste0("./analysis/tSNE/leu/",i,"/tSNE_LeuLin.eps")) + postscript(paste0("./analysis/tSNE/leu/",i,"/tSNE_res",opt$res.leu,".eps")) plot <- TSNEPlot(object=sc10x.Leu,pt.size=5,do.label=TRUE,label.size=10,do.return=TRUE,vector.friendly=FALSE) plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) @@ -296,43 +311,142 @@ for (i in c("Pb","Pc","Pd")){ dev.off() rm(plot) - sc10x <- scMerge(sc10x,sc10x.Epi,sc10x.St,i.1="Epi.dws.sc",i.2="St.dws.sc",nm="Merge") + genes.leu <- read_excel("genesets/40425_2017_215_MOESM1_ESM.xlsx",sheet="S3. Candidate markers") + leu <- as.list(unique(genes.leu[,2]))$Cell + leu.l <- leu[-c(1,3,4,7:9,14,15,17:18,20:21,23:30)] + leu.t <- leu[-c(1,2,5,6,8:13,17:20,22)] +# genes.leu.s <- genes.leu[genes.leu$Selected==1,] +# leu.s <- as.list(unique(genes.leu.s[,2]))$Cell + genes.leu <- genes.leu[unlist(genes.leu[,2]) %in% unlist(leu),] + genes.leu.l <- genes.leu[unlist(genes.leu[,2]) %in% unlist(leu.l),] +# genes.leu.s <- genes.leu.s[unlist(genes.leu.s[,2]) %in% unlist(leu),] +genes.leu.t <- genes.leu[unlist(genes.leu[,2]) %in% unlist(leu.t),] - sc10x@ident <- factor(sc10x@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) - sctSNE3CustCol(sc10x,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=i) - - if (i=="Pd"){ - for (j in list(c("D17PrPzF_Via","D27PrPzF_Via","D35PrPzF_Via"),c("D17PrTzF_Via","D27PrTzF_Via","D35PrTzF_Via"))){ - sc10x.sub <- scSubset(sc10x,"samples",j) - sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") - sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) - sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".",substr(j[1],6,7))) - } - rm(sc10x.sub) - } else if (i=="Pb"){ - sc10x.sub <- scSubset(sc10x,"Glandular","Glandular") - sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") - sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) - sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".Glandular")) - - sc10x.sub <- scSubset(sc10x,"Stromal","Stromal") - sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") - sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) - sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".Stromal")) +# gene.set1 <- split(genes.leu[,1], genes.leu[,2]) +# gene.set1 <- lapply(gene.set1,unname) +# gene.set1 <- lapply(gene.set1,unlist) +# gene.set <- gene.set1 +# rm(gene.set1) +# gc() +# min.leu <- min(table(sc10x.Leu@meta.data[,paste0("res",opt$res.leu)])) +# results <- scQuSAGE(sc10x.Leu,gs=gene.set,res.use=opt$res.leu,ds=0,nm="Leu",folder=paste0("leu/",i)) +# sc10x.Leu <- results[[1]] +# results.cor.Leu <- results[[2]] +# results.clust.Leu.id <- results[[3]] +# rm(results) +# rm(gene.set) +# gene.set1 <- split(genes.leu.s[,1], genes.leu.s[,2]) +# gene.set1 <- lapply(gene.set1,unname) +# gene.set1 <- lapply(gene.set1,unlist) +# gene.set <- gene.set1 +# rm(gene.set1) +# gc() +# sc10x.Leu <- SetAllIdent(object=sc10x.Leu,id=paste0("res",opt$res.leu)) +# results <- scQuSAGE(sc10x.Leu,gs=gene.set,res.use=opt$res.leu,ds=0,nm="Leu.s",folder=paste0("leu/",i)) +# sc10x.Leu <- results[[1]] +# results.cor.Leu.s <- results[[2]] +# results.clust.Leu.s.id <- results[[3]] +# rm(results) +# rm(gene.set) + gene.set1 <- split(genes.leu.l[,1], genes.leu.l[,2]) + gene.set1 <- lapply(gene.set1,unname) + gene.set1 <- lapply(gene.set1,unlist) + gene.set <- gene.set1 + rm(gene.set1) + gc() + sc10x.Leu <- SetAllIdent(object=sc10x.Leu,id=paste0("res",opt$res.leu)) + results <- scQuSAGE(sc10x.Leu,gs=gene.set,res.use=opt$res.leu,ds=0,nm="Leu.l",folder=paste0("leu/",i)) + sc10x.Leu <- results[[1]] + results.cor.Leu.l <- results[[2]] + results.clust.Leu.l.id <- results[[3]] + rm(results) + rm(gene.set) - for (j in c("BPH327PrGF_Via","BPH327PrSF_Via")){ - sc10x.sub <- scSubset(sc10x,"samples",j) - sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") - sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) - sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".",substr(j,4,6),substr(j,9,9))) - } - } + sc10x.T <- scSubset(sc10x.Leu,i="Leu.l",g="T-cells") - write.table(table(sc10x@meta.data[,"samples"],sc10x@meta.data[,"Merge"]),file=paste0("./analysis/table/Table_samples_Merge",i,".csv"),row.names=TRUE,col.names=NA,append=FALSE,sep=",") - write.table(round(prop.table(table(sc10x@meta.data[,"samples"],sc10x@meta.data[,"Merge"]),1)*100,1),file=paste0("./analysis/table/ProbTable_samples_Merge",i,".csv"),row.names=TRUE,col.names=NA,append=FALSE,sep=",") + sc10x.T <- scCluster(sc10x.T,pc.use=opt$acca,res.use=opt$res.leu,folder=paste0("t/",i),red="cca.aligned") + sc10x.T <- StashIdent(object=sc10x.T,save.name=paste0("res",opt$res.leu)) + sc10x.T <- RunTSNE(object=sc10x.T,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) + postscript(paste0("./analysis/tSNE/t/",i,"/tSNE_Sample.eps")) + plot <- TSNEPlot(object=sc10x.T,group.by="samples",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() + postscript(paste0("./analysis/tSNE/t/",i,"/tSNE_res",opt$res.leu,".eps")) + plot <- TSNEPlot(object=sc10x.T,pt.size=5,do.label=TRUE,label.size=10,do.return=TRUE,vector.friendly=FALSE) + plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) + plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) + plot(plot) + dev.off() + rm(plot) + + gene.set1 <- split(genes.leu.t[,1], genes.leu.t[,2]) + gene.set1 <- lapply(gene.set1,unname) + gene.set1 <- lapply(gene.set1,unlist) + gene.set <- gene.set1 + rm(gene.set1) + gc() + sc10x.T <- SetAllIdent(object=sc10x.T,id=paste0("res",opt$res.leu)) + results <- scQuSAGE(sc10x.T,gs=gene.set,res.use=opt$res.leu,ds=0,nm="Leu.t",folder=paste0("t/",i)) + sc10x.Leu <- results[[1]] + results.cor.Leu.l <- results[[2]] + results.clust.Leu.l.id <- results[[3]] + rm(results) + rm(gene.set) + + + +# +# sc10x.Merge <- scMerge(sc10x.nEpi,sc10x.St,sc10x.Leu,i.1="St.dws.sc",i.2="Leu",nm="Merge") +# sc10x.Merge <- scMerge(sc10x,sc10x.Epi,sc10x.Merge,i.1="Epi.dws.sc",i.2="Merge",nm="Merge") +# +# sc10x.Merge@ident <- factor(sc10x.Merge@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo",leu)) +# +# postscript(paste0("./analysis/tSNE/FINAL/tSNE_Merge_",i,".eps")) +# plot <- TSNEPlot(object=sc10x.Merge,pt.size=2.5,do.label=TRUE,label.size=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() +# rm(plot) +# +# sctSNE3CustCol(sc10x,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=i) +# +# if (i=="Pd"){ +# for (j in list(c("D17PrPzF_Via","D27PrPzF_Via","D35PrPzF_Via"),c("D17PrTzF_Via","D27PrTzF_Via","D35PrTzF_Via"))){ +# sc10x.sub <- scSubset(sc10x,"samples",j) +# sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") +# sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) +# sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".",substr(j[1],6,7))) +# } +# rm(sc10x.sub) +# } else if (i=="Pb"){ +# sc10x.sub <- scSubset(sc10x,"Glandular","Glandular") +# sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") +# sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) +# sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".Glandular")) +# +# sc10x.sub <- scSubset(sc10x,"Stromal","Stromal") +# sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") +# sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) +# sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".Stromal")) +# +# for (j in c("BPH327PrGF_Via","BPH327PrSF_Via")){ +# sc10x.sub <- scSubset(sc10x,"samples",j) +# sc10x.sub <- SetAllIdent(object=sc10x.sub,id="Merge") +# sc10x.sub@ident <- factor(sc10x.sub@ident,levels=c("BE","LE","Hillock","Club","Fib","SM","Endo","B-cells","T-cells","Macrophages","Mast cells")) +# sctSNE3CustCol(sc10x.sub,i="Merge",bl=c("BE","LE","Hillock","Club"),rd=c("Fib","SM","Endo"),gn=c("B-cells","T-cells","Macrophages","Mast cells"),file=paste0(i,".",substr(j,4,6),substr(j,9,9))) +# } +# } +# +# write.table(table(sc10x@meta.data[,"samples"],sc10x@meta.data[,"Merge"]),file=paste0("./analysis/table/Table_samples_Merge",i,".csv"),row.names=TRUE,col.names=NA,append=FALSE,sep=",") +# write.table(round(prop.table(table(sc10x@meta.data[,"samples"],sc10x@meta.data[,"Merge"]),1)*100,1),file=paste0("./analysis/table/ProbTable_samples_Merge",i,".csv"),row.names=TRUE,col.names=NA,append=FALSE,sep=",") + assign(paste0("sc10x.",i,".analized"),sc10x) assign(paste0("sc10x.Epi.",i,".analized"),sc10x.Epi) + assign(paste0("sc10x.nEpi.",i,".analized"),sc10x.nEpi) assign(paste0("sc10x.St.",i,".analized"),sc10x.St) assign(paste0("sc10x.Leu.",i,".analized"),sc10x.Leu) assign(paste0("results.cor.Lin.",i),results.cor.Lin) @@ -341,6 +455,13 @@ for (i in c("Pb","Pc","Pd")){ assign(paste0("results.clust.Epi.id.",i),results.clust.Epi.id) assign(paste0("results.cor.St.",i),results.cor.St) assign(paste0("results.clust.St.id.",i),results.clust.St.id) + assign(paste0("results.cor.Leu.",i),results.cor.Leu) + assign(paste0("results.clust.Leu.id.",i),results.clust.Leu.id) + assign(paste0("results.cor.Leu.s.",i),results.cor.Leu.s) + assign(paste0("results.clust.Leu.s.id.",i),results.clust.Leu.s.id) + assign(paste0("results.cor.Leu.l.",i),results.cor.Leu.l) + assign(paste0("results.clust.Leu.l.id.",i),results.clust.Leu.l.id) + rm(sc10x.sub) rm(sc10x) rm(sc10x.Epi) @@ -349,13 +470,21 @@ for (i in c("Pb","Pc","Pd")){ rm(results.cor.Lin) rm(results.cor.Epi) rm(results.cor.St) + rm(results.cor.Leu) + rm(results.cor.Leu.s) + rm(results.cor.Leu.l) rm(results.clust.Lin.id) rm(results.clust.Epi.id) rm(results.clust.St.id) + rm(results.clust.Leu.id) + rm(results.clust.Leu.s.id) + rm(results.clust.Leu.l.id) rm(min.all) rm(min.epi) + rm(min.nepi) rm(min.st) - rm(i) + rm(min.leu) } +rm(i) save.image(file="./analysis/sc10x.analized.RData")