Effect of Srsf2 P95H mutation on RNA splicing and expression in vivo. (A) Number of abnormal AS events identified in KSL cells of KI mice compared with that of WT mice in transplantation settings (n = 6 pairs). The log₁₀ ratio of the number of abnormal AS events differentially included to those differentially excluded is also shown. (B) Inclusion levels of the CEs and mutually exclusive exons in KSL cells of KI mice compared with that of WT mice in transplantation settings are plotted. The colored dots represent significantly more included (red) and excluded (blue) events. The gray dots represent nonsignificant AS events with ≤5% inclusion level differences. (C) The composition of SSNG motifs in the more included or excluded CEs in the KSL cells of KI mice compared with that of WT mice in transplantation settings, and SSNG motif compositions in all of the exons from the mouse reference genome (mm10). (D) Relative frequency (log₁₀ ratio) of the 7-base window-averaged number of CCNG and GGNG motifs in the CEs promoted vs repressed in KSL cells of KI mice, extending 100 nt into their upstream and downstream from the 5′ and 3′ splice sites. (E) Venn diagram comparing the differentially spliced genes between KI and WT cells from 4 different populations, ie, KSL and MP cells from the steady-state mice and those from the recipient mice in transplantation settings. Genes registered in the Cancer Gene Census are shown in red. (F) MA plot and volcano plot showing the transcriptional changes between KI and WT KSL cells in transplantation settings. Significantly upregulated and downregulated genes in KI cells compared with control cells are indicated by red and blue, respectively. (G) Venn diagram comparing the differentially upregulated (left) and downregulated (right) genes in Srsf2 mutant KSL and MP cells from the steady-state mice vs those from the recipient mice in transplantation settings. Genes registered in the Cancer Gene Census are shown in red. (H) Gene set enrichment analysis demonstrating a significant positive enrichment of gene sets upregulated in human MDS compared with healthy controls in Srsf2 mutant KSL cells relative to wild-type KSL cells in transplantation settings (left panel), and negative enrichment of gene sets associated with the stemness (middle panel) and quiescence (right panel) in Srsf2 mutant KSL cells relative to wild-type KSL cells under the steady-state conditions. Nominal P values, FDRs, and normalized enrichment scores (NSEs) are indicated. KI, Srsf2 mutant transplanted mice; WT, wild-type transplanted mice.