Entering edit mode
15 months ago
Ann
▴
40
I would like to use LogFC aftre RdgeR to colour KEGG Pathway Maps with my data.
- Is it ok to extract $logFC, create a named list and put it into pathway()?
- Should I use some filters like FDR < 0.05 & LogFC > or < 1.5 before using pathview()? I want to get grey colour for all NoSignal, red for Up and blue for Down regulated genes
Picture I want to get:
My EdgeR pipeline:
# filtering
keep <- filterByExpr(y, design)
y_filtered <- y[keep, , keep.lib.sizes=FALSE]
# to estimate dispersion
y_disp_design <- estimateDisp(y_filtered, design = design, verbose=TRUE)
# Fit a quasi-likelihood negative binomial generalized log-linear model to count data
fit <- glmQLFit(y_disp_design, design, robust = TRUE)
# Conduct genewise statistical tests
LFC <- 1.5
tr.c1_2 <- glmTreat (fit, contrast = my.contrasts[, 'c1_2'], lfc = log2(LFC))
# Identify which genes are significantly differentially expressed
is.de.tr.c1_2 <- decideTestsDGE(tr.c1_2)
# View the amount of Up and Down genes
summary(is.de.tr.c1_2)
Down 3191
NotSig 13930
Up 1077
# Extracts the most differentially expressed genes
TT_tr.c1_2 <- topTags(tr.c1_2, Inf)
# View TT_tr.c1_2
TT_tr.c1_2
Coefficient: 1*stage_1 -1*stage_2
logFC unshrunk.logFC logCPM PValue FDR
TRINITY_DN583_c0_g1 -3.254507 -3.255309e+00 6.980710 1.123231e-11 2.044055e-07
TRINITY_DN1301_c4_g1 -5.887508 -5.921260e+00 3.852718 2.272249e-11 2.067519e-07
...
# extract LogFC !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
gene_list <- TT_tr.c1_2$table$logFC
# create named list
names(gene_list) <- my_KO_names
# view named logFC
head(gene_list)
K14156 K10860 K00072 K15701 K22257 K06240
-3.960318 -3.503302 -3.774908 -3.335891 -4.597206 -3.679689
dme <- pathview(gene.data=gene_list,
pathway.id="04064",
species = "ko",
out.suffix = my_pair,
limit = list(gene = 2,
cpd = 1))