Figure 1
Transcriptome-wide analysis of the stability of candidate reference genes for quantitative RT-PCR assays across six life-cycle stages of S. mansoni. Twenty-four RNA-Seq libraries comprising six S. mansoni life-cycle stages were analyzed using four different normalization methods, namely DESeq2, TMM.CPM, UQ, and TPM. (A) Box plots of the coefficients of variation (CV), which were calculated based on the expression levels of the 272 genes with the lowest coefficients of variation, representing 2% of all genes detected in the RNA-Seq libraries, and analyzed with each of the four normalization methods indicated in the x-axis. The horizontal line represents the CVs’ median for each normalization method, while the + signal represents the mean. The boxes and whiskers represent the inter-quantile and min to max ranges, respectively. (B) Venn Diagram shows the number of genes and the overlap among the 272 most stable genes found in each of the four RNA-Seq normalization methods. (C) Venn Diagram showing the number of genes and the overlap among the 272 most stable genes in the DESeq2, TMM.CPM and UQ normalization methods. (D) Venn Diagram showing the number of genes and overlap among the ten most stable genes according to their CVs in each RNA-Seq normalization method tested. UQ upper quartile, TMM trimmed mean of M-values, CPM counts per million, TPM transcripts per million.
