Question: Tukeys'S Hsd After Anova Using R
2
gravatar for deschisandy
6.0 years ago by
deschisandy60
deschisandy60 wrote:
The dataset has 1000 genes and contains 24 samples with two mouse strains tested (129 and B6) and six brain regions. There are two replicates for each region.

The ANOVA was performed as follows:

      sdata<-read.table("http://www.chibi.ubc.ca/wp-content/uploads/2013/02/
                sandberg-sampledata.txt", header=T, row.names=1)
     strain <- gl(2,12,24, label=c("129","bl6"))
     region <- gl(6,2,24, label=c("ag", "cb", "cx", "ec", "hp", "mb"))
   # define ANOVA function ##  ANOVA
   aof <- function(x) { 
   m<-data.frame(strain,region, x); 
   anova(aov(x ~ strain + region + strain*region, m))
   }
 # apply analysis to the data and get the pvalues.
 anovaresults <- apply(sdata, 1, aof)

pvalues<-data.frame( lapply(anovaresults, function(x) { x["Pr(>F)"][1:3,] })    )

 # Get the genes with good region effect pvalues. 
 reg.hi.p <-t(data.frame(pvalues[2, pvalues[2,] < 0.0001 & pvalues[3,] > 0.1]))
 reg.hi.pdata <- sdata[ row.names(reg.hi.p), ]

A significant p-value resulting from a 1-way ANOVA test would indicate that a gene is differentially expressed in at least one of the groups analyzed. Now that there are more than two groups being analyzed, however, the 1-way ANOVA does not specifically indicate which pair of groups exhibits statistical differences. I know that Post Hoc tests can be applied in this specific situation to determine which specific pair/pairs are differentially expressed in each of the regions ( irrespective of the strains). I would like to know how to apply the Tukey's HSD using R in this case to find out which of these genes ( the ones with good region effect pvalues) are expressed in which region ( for instance in which brain region like "ag","cb","cx and so on).

I tried doing it this way but generates the following error:

                  mcps <- TukeyHSD(anovaresults, ordered=TRUE) 
                  Error in UseMethod("TukeyHSD") :  no applicable method for 'TukeyHSD' applied to an object of class "list"
R differential-expression • 3.7k views
ADD COMMENTlink modified 6.0 years ago • written 6.0 years ago by deschisandy60
2
gravatar for David W
6.0 years ago by
David W4.7k
New Zealand
David W4.7k wrote:

see ?TukeyHSD. The function can take an anova fit (as returned by aov) but not a list or a an ANOVA table which is what you have in your list. You'll need to edit the function you're using to run your ANOVAs so that they return the aov object.

Note though, a p-value < 0.05 doesn't guarantee you'll have significant pairwise-comparisons. Also, these aren't "one way" ANOVAs -- you have two predictor-variables.

ADD COMMENTlink modified 6.0 years ago • written 6.0 years ago by David W4.7k

I noticed that it is a list object but am unsure how to create a anova fit now.

ADD REPLYlink written 6.0 years ago by deschisandy60

Just edited the answer to make it more clear (and point out these are not "one way ANOVAs")

ADD REPLYlink modified 6.0 years ago • written 6.0 years ago by David W4.7k
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