Global gene expression is dysregulated by single and double mutants. (A) Unsupervised principal component analysis of differentially expressed genes in murine LK cells of each genotype (WT, n = 3; Srsf2^P95H/+, n = 2; Runx1 knockout, n = 3; double mutant, n = 3). (B) Venn diagram showing the overlap between differentially expressed genes of each genotype compared with WT murine LK cells. (C) Unsupervised principal component analysis of differentially expressed genes in K562 cells of each genotype (n = 3 for each genotype). (D) Venn diagram showing the overlap between differentially expressed genes of each genotype compared with WT K562 cells. (E) GO enrichment analysis of the downregulated genes in double mutant K562 cells compared with WT (left) and in double mutant murine LK cells compared with WT (right). (F) GO enrichment analysis of the upregulated genes in double mutant K562 cells compared with WT (left) and in double mutant murine LK cells compared with WT (right). ClusterProfiler³³ was used to refine overlapping GOs by calculating enriched functional categories of individual gene clusters for panels E and F. The colors of the circles indicate the P values and the sizes of the circles indicate the number of genes in each GO term. (G) Growth curves of single/double mutant K562 cells. Cells were seeded in triplicate on day 1 and counted daily using trypan blue exclusion. Data are mean ± standard deviation of 3 independent experiments. ∗P < .05. (H) Percentages of early apoptotic cells (annexinV⁺ 7AAD⁻) in single/double mutant K562 cells were analyzed by flow cytometry on day 6 and 9 after short hairpin RNA transduction and puromycin selection. Cells were seeded in duplicate in 3 independent experiments. ∗P < .05, ∗∗P < .01. GO, gene ontology; KD, knockdown.