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## NASH liver GPL15207 Affymetrix Human Gene Expression Array | ||
## Patients were grouped according histology: | ||
## group I (<5% steatosis), group II (NAFLD, 30-50% steatosis) and group III (NASH). | ||
rm(list=ls()) | ||
library(GEOquery) | ||
library(limma) | ||
GSE59045 <- getGEO('GSE59045', destdir=".",getGPL = F) | ||
exprSet=exprs(GSE59045[[1]]) | ||
library("annotate") | ||
GSE59045[[1]] | ||
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pdata=pData(GSE59045[[1]]) | ||
treatment=factor(c(rep('steatosis',6),rep('NAFLD',4),rep('NASH',5)) ) | ||
treatment=relevel(treatment,'steatosis') | ||
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#platformDB='hgu133plus2.db' | ||
#library(platformDB, character.only=TRUE) | ||
probeset <- featureNames(GSE59045[[1]]) | ||
library(Biobase) | ||
library(GEOquery) | ||
#Download GPL file, put it in the current directory, and load it: | ||
gpl <- getGEO('GPL15207', destdir=".") | ||
colnames(Table(gpl)) ## [1] 41108 17 | ||
head(Table(gpl)[,c(1,15,19)]) ## you need to check this , which column do you need | ||
probe2symbol=Table(gpl)[,c(1,15)] | ||
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symbol=probe2symbol[match(probeset,probe2symbol[,1]),2] | ||
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a=cbind(symbol,exprSet) | ||
## remove the duplicated probeset | ||
rmDupID <-function(a=matrix(c(1,1:5,2,2:6,2,3:7),ncol=6)){ | ||
exprSet=a[,-1] | ||
rowMeans=apply(exprSet,1,function(x) mean(as.numeric(x),na.rm=T)) | ||
a=a[order(rowMeans,decreasing=T),] | ||
exprSet=a[!duplicated(a[,1]),] | ||
# | ||
exprSet=exprSet[!is.na(exprSet[,1]),] | ||
rownames(exprSet)=exprSet[,1] | ||
exprSet=exprSet[,-1] | ||
return(exprSet) | ||
} | ||
exprSet=rmDupID(a) | ||
rn=rownames(exprSet) | ||
exprSet=apply(exprSet,2,as.numeric) | ||
rownames(exprSet)=rn | ||
exprSet[1:4,1:4] | ||
#exprSet=log(exprSet) ## based on e | ||
boxplot(exprSet,las=2) | ||
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design=model.matrix(~ treatment) | ||
fit=lmFit(exprSet,design) | ||
fit=eBayes(fit) | ||
vennDiagram(decideTests(fit)) | ||
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write.csv(topTable(fit,coef='treatmentNASH',n=Inf,adjust='BH'),"NASH-steatosis.DEG.csv") | ||
write.csv(topTable(fit,coef='treatmentNAFLD',n=Inf,adjust='BH'),"NAFLD-steatosis.DEG.csv") |