test_that("LFC shrinkage works", {
  # testing out various methods for LFC shrinkage
  dds <- makeExampleDESeqDataSet(betaSD=1,n=1000,m=20)
  expect_error(lfcShrink(dds, coef=2), "first run")
  dds <- DESeq(dds)
  res <- results(dds, name="condition_B_vs_A")

  # dds and res must match
  expect_error(lfcShrink(dds=dds, coef=2, res=res[1:500,], type="normal"), "rownames")
  expect_error(lfcShrink(dds=dds, coef=2, res=res[1:500,], type="apeglm"), "rownames")

  # some quick contrast tests
  expect_error(lfcShrink(dds=dds, contrast=c("treatment","B","A"), res=res, type="normal"))
  expect_error(lfcShrink(dds=dds, contrast=c("condition","C","A"), res=res, type="normal"))

  # try out various types and ways of specifying coefs
  res.n <- lfcShrink(dds=dds, coef="condition_B_vs_A", res=res, type="normal")
  res.n <- lfcShrink(dds=dds, coef=2, res=res, type="normal")
  res.n <- lfcShrink(dds=dds, coef=2, type="normal")
  res.ape <- lfcShrink(dds=dds, coef=2, type="apeglm")
  res.ash <- lfcShrink(dds=dds, res=res, type="ashr")

  # prior info
  # ranged versions
  gr.res <- results(dds, name="condition_B_vs_A", format="GRanges")
  expect_error(lfcShrink(dds=dds, coef=2, type="normal", res=gr.res), "GRanges")
  gr.res <- lfcShrink(dds=dds, coef=2, type="normal", format="GRanges")
  gr.res <- lfcShrink(dds=dds, coef=2, type="apeglm", format="GRanges")
  gr.res <- lfcShrink(dds=dds, coef=2, type="ashr", format="GRanges")
  priorInfo(mcols(gr.res)) # still has priorInfo() on the metadata columns

  # plot against true
  ## par(mfrow=c(1,3))
  ## plot(mcols(dds)$trueBeta, res.n$log2FoldChange); abline(0,1,col="red")
  ## plot(mcols(dds)$trueBeta, res.ape$log2FoldChange); abline(0,1,col="red")
  ## plot(mcols(dds)$trueBeta, res.ash$log2FoldChange); abline(0,1,col="red")

  # LFC threshold for "normal" and "apeglm"
  res0 <- results(dds, name="condition_B_vs_A", lfcThreshold=1)
  res.n <- lfcShrink(dds=dds, coef=2, type="normal", lfcThreshold=1)
  res.ape <- lfcShrink(dds=dds, coef=2, type="apeglm", lfcThreshold=1)

  # this doesn't work in R-devel as of June 2019...
  #summary.res <- capture.output({ summary(res.n) })
  #expect_true(any(grepl("1.00", summary.res)))
  #plotMA(res0, ylim=c(-4,4), cex=1); abline(h=c(-1,1),col="dodgerblue")
  #plotMA(res.n, ylim=c(-4,4), cex=1); abline(h=c(-1,1),col="dodgerblue")
  #plotMA(res.ape, ylim=c(-4,4), cex=1); abline(h=c(-1,1),col="dodgerblue")
  # s-value returned
  res.ape <- lfcShrink(dds=dds, coef=2, type="apeglm", svalue=TRUE)
  expect_true("svalue" %in% names(res.ape))
  res.ash <- lfcShrink(dds=dds, res=res, type="ashr", svalue=TRUE)
  expect_true("svalue" %in% names(res.ash))

  # plotMA works with s-values
  plotMA(res.ape, cex=1)
  plotMA(res.ash, cex=1)

  # summary works with s-values
  summary.res <- capture.output({ summary(res.ape) })
  summary.res <- capture.output({ summary(res.ash) })
  # list returned
  res.ape <- lfcShrink(dds=dds, coef=2, type="apeglm", returnList=TRUE)
  res.ash <- lfcShrink(dds=dds, res=res, type="ashr", returnList=TRUE)

  # test wrong coef specified
  resInt <- results(dds, name="Intercept")
  expect_error(lfcShrink(dds=dds, coef=2, res=resInt, type="apeglm"))

  # test supplied model.matrix
  full <- model.matrix(~condition, colData(dds))
  dds <- DESeq(dds, full=full)
  res <- results(dds)
  res.normal <- lfcShrink(dds=dds, coef=2, res=res, type="normal")
  res.ape <- lfcShrink(dds=dds, coef=2, res=res, type="apeglm")

  # only running LRT upstream
  dds <- makeExampleDESeqDataSet(m=4)
  mm <- model.matrix(~condition, colData(dds))
  mm0 <- model.matrix(~1, colData(dds))
  dds <- DESeq(dds, full=mm, reduced=mm0, test="LRT")
  res.normal <- lfcShrink(dds, coef="conditionB", type="normal")

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DESeq2 documentation built on Feb. 22, 2021, 10 a.m.