Combined impact of mutations in SF3B1 and inv(3)/t(3;3) on gene expression and RNA splicing. (A) Similarity matrix and hierarchical clustering of 4 groups (Mx1-Cre control, Mx1-Cre inv(3), Mx1-Cre Sf3b1^K700E/WT, and Mx1-Cre inv(3) Sf3b1^K700E/WT) by differential gene expression. Three samples were independently collected in each group. (B) Principal component (PC) analysis of gene expression from 12 samples (4 groups, biologically triplicated). (C) Overlap of differentially expressed genes compared with Mx1-Cre control. (D) Significantly dysregulated pathways. P values are color-coded. (E) Significantly dysregulated pathways. Number of genes and statistical significance (−log₁₀FDR) were shown. The impact of SF3B1 mutation on gene expression was analyzed under the condition with or without EVI1 rearrangement. (F) Overlap of differentially spliced genes compared with AML without SF3B1 mutation or EVI1 rearrangement. (G) Aberrant splicing detected in AML with EVI1 rearrangement and SF3B1 mutations. x-axis and y-axis indicate the percent spliced in (ψ) of each splicing event in the presence/absence of genetic alterations. Alternate splice sites, mutually exclusive exons, retained introns, or cassette exons are shown when P < .01. Red and blue dots represent individual splicing events or coding genes that are promoted or repressed in each condition; green dots are shown when the difference in percent spliced is <10%. The number of aberrantly spliced genes is indicated in blue or red.