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basic visualization for expression matrix
jmzeng1314@163.com
March 14, 2017
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安装并加载必须的packages
代码语言:javascript复制如果你还没有安装,就运行下面的代码安装:
BiocInstaller::biocLite('CLL')install.packages('corrplot')install.packages('gpairs')install.packages('vioplot')
代码语言:javascript复制如果你安装好了,就直接加载它们即可
library(CLL)library(ggplot2)library(reshape2)library(gpairs)library(corrplot)
加载内置的测试数据:
代码语言:javascript复制data(sCLLex)sCLLex=sCLLex[,1:8] ## 样本太多,我就取前面8个
group_list=sCLLex$DiseaseexprSet=exprs(sCLLex)head(exprSet)
代码语言:javascript复制## CLL11.CEL CLL12.CEL CLL13.CEL CLL14.CEL CLL15.CEL CLL16.CEL
## 1000_at 5.743132 6.219412 5.523328 5.340477 5.229904 4.920686
## 1001_at 2.285143 2.291229 2.287986 2.295313 2.662170 2.278040
## 1002_f_at 3.309294 3.318466 3.354423 3.327130 3.365113 3.568353
## 1003_s_at 1.085264 1.117288 1.084010 1.103217 1.074243 1.073097
## 1004_at 7.544884 7.671801 7.474025 7.152482 6.902932 7.368660
## 1005_at 5.083793 7.610593 7.631311 6.518594 5.059087 4.855161
## CLL17.CEL CLL18.CEL
## 1000_at 5.325348 4.826131
## 1001_at 2.350796 2.325163
## 1002_f_at 3.502440 3.394410
## 1003_s_at 1.091264 1.076470
## 1004_at 6.456285 6.824862
## 1005_at 5.176975 4.874563
代码语言:javascript复制group_list
代码语言:javascript复制## [1] progres. stable progres. progres. progres. progres. stable stable
## Levels: progres. stable
接下来进行一系列绘图操作
代码语言:javascript复制主要用到ggplot2这个包,需要把我们的宽矩阵用reshape2包变成长矩阵
library(reshape2)exprSet_L=melt(exprSet)colnames(exprSet_L)=c('probe','sample','value')exprSet_L$group=rep(group_list,each=nrow(exprSet))head(exprSet_L)
代码语言:javascript复制## probe sample value group
## 1 1000_at CLL11.CEL 5.743132 progres.
## 2 1001_at CLL11.CEL 2.285143 progres.
## 3 1002_f_at CLL11.CEL 3.309294 progres.
## 4 1003_s_at CLL11.CEL 1.085264 progres.
## 5 1004_at CLL11.CEL 7.544884 progres.
## 6 1005_at CLL11.CEL 5.083793 progres.
boxplot
代码语言:javascript复制p=ggplot(exprSet_L,aes(x=sample,y=value,fill=group)) geom_boxplot()print(p)
vioplot
代码语言:javascript复制#library(vioplot)#?vioplot#vioplot(exprSet)#do.call(vioplot,c(unname(exprSet),col='red',drawRect=FALSE,names=list(names(exprSet))))p=ggplot(exprSet_L,aes(x=sample,y=value,fill=group)) geom_violin()print(p)
histogram
代码语言:javascript复制p=ggplot(exprSet_L,aes(value,fill=group)) geom_histogram(bins = 200) facet_wrap(~sample, nrow = 4)print(p)
density
代码语言:javascript复制p=ggplot(exprSet_L,aes(value,col=group)) geom_density() facet_wrap(~sample, nrow = 4)print(p)
代码语言:javascript复制p=ggplot(exprSet_L,aes(value,col=group)) geom_density() print(p)
gpairs
代码语言:javascript复制library(gpairs)gpairs(exprSet
#,upper.pars = list(scatter = 'stats')
#,lower.pars = list(scatter = 'corrgram')
)
cluster
代码语言:javascript复制out.dist=dist(t(exprSet),method='euclidean')out.hclust=hclust(out.dist,method='complete')plot(out.hclust)
PCA
代码语言:javascript复制pc <- prcomp(t(exprSet),scale=TRUE)pcx=data.frame(pc$x)pcr=cbind(samples=rownames(pcx),group_list, pcx) p=ggplot(pcr, aes(PC1, PC2)) geom_point(size=5, aes(color=group_list))
geom_text(aes(label=samples),hjust=-0.1, vjust=-0.3)print(p)
heatmap
代码语言:javascript复制choose_gene=names(sort(apply(exprSet, 1, mad),decreasing = T)[1:50])choose_matrix=exprSet[choose_gene,]choose_matrix=scale(choose_matrix)heatmap(choose_matrix)
代码语言:javascript复制library(gplots)
代码语言:javascript复制##
## Attaching package: 'gplots'
代码语言:javascript复制## The following object is masked from 'package:stats':
##
## lowess
代码语言:javascript复制heatmap.2(choose_matrix)
代码语言:javascript复制library(pheatmap)pheatmap(choose_matrix)
DEG && volcano plot
代码语言:javascript复制library(limma)
代码语言:javascript复制##
## Attaching package: 'limma'
代码语言:javascript复制## The following object is masked from 'package:BiocGenerics':
##
## plotMA
代码语言:javascript复制design=model.matrix(~factor(group_list))fit=lmFit(exprSet,design)fit=eBayes(fit)DEG=topTable(fit,coef=2,n=Inf)with(DEG, plot(logFC, -log10(P.Value), pch=20, main="Volcano plot"))
代码语言:javascript复制logFC_cutoff <- with(DEG,mean(abs( logFC)) 2*sd(abs( logFC)) )DEG$change = as.factor(ifelse(DEG$P.Value < 0.05 & abs(DEG$logFC) > logFC_cutoff, ifelse(DEG$logFC > logFC_cutoff ,'UP','DOWN'),'NOT')
)this_tile <- paste0('Cutoff for logFC is ',round(logFC_cutoff,3), 'nThe number of up gene is ',nrow(DEG[DEG$change =='UP',]) , 'nThe number of down gene is ',nrow(DEG[DEG$change =='DOWN',]))g = ggplot(data=DEG, aes(x=logFC, y=-log10(P.Value), color=change))
geom_point(alpha=0.4, size=1.75)
theme_set(theme_set(theme_bw(base_size=20)))
xlab("log2 fold change") ylab("-log10 p-value")
ggtitle( this_tile ) theme(plot.title = element_text(size=15,hjust = 0.5))
scale_colour_manual(values = c('blue','black','red')) ## corresponding to the levels(res$change)print(g)
ggplot画图是可以切换主题的
代码语言:javascript复制其实绘图有非常多的细节可以调整,还是略微有点繁琐的!
p=ggplot(exprSet_L,aes(x=sample,y=value,fill=group)) geom_boxplot()print(p)
代码语言:javascript复制p=p stat_summary(fun.y="mean",geom="point",shape=23,size=3,fill="red")p=p theme_set(theme_set(theme_bw(base_size=20)))p=p theme(text=element_text(face='bold'),axis.text.x=element_text(angle=30,hjust=1),axis.title=element_blank())print(p)
可以很明显看到,换了主题之后的图美观一些。