The Integrator complex regulates differential snRNA processing and fate of adult stem cells in the highly regenerative planarian Schmidtea mediterranea
Fig 5
ints RNAi interferes with histone mRNA processing and disrupts the splicing- and gene expression profiles of neoblasts.
(A) qRT-PCR quantification showing that the 3’-unprocessed mRNAs of H2A, H3 and H4 accumulate in regenerating ints RNAi animals (3 dpa; 16 days of RNAi). Expression levels are the averages of three biological replicates normalized to those of the total pool of the respective histone (Error bars represent standard deviation; two-sided t-test, * p<0.05, ** p<0.01). (B) Overview of the splicing events significantly changed in neoblasts depleted of ints3, ints9 or in both conditions showing that the exclusion of alternative (Alt.) exons (exon skipping) is the primary splicing defect induced by ints RNAi. PSI = Percent Spliced-In. For details see S4 Table. (C) Two examples of ints RNAi-induced exon skipping events in neoblasts affecting transcripts encoding putative homologs of dspp and espl1, analyzed by RT-PCR. Results for all other RT-PCR validations are shown in S7A Fig. (D) Comparison of the gene expression changes induced by ints RNAi in X1 cells to the transcripts differentially expressed between the X1, X2 and Xin FACS fractions [41]. Transcripts annotated as stem cell-specific were statistically significantly over-represented among the transcripts down-regulated in ints RNAi X1 cells (threshold for differentially expressed genes in ints RNAi neoblasts: padj<0.05; |log2 fold-change| > 1; Statistical test for over-/under-representation: hypergeometric test, * p<0.05, *** p<0.001).