Kmt2d and Trp53 deficiencies coordinately induce B-cell lymphoma with Yap1 overexpression. (A) Survival curves of recipient mice transplanted with mock (n = 7), sgKmt2d (n = 27), sgTrp53 (n = 9), and double-targeted (n = 9) HSPCs with constitutive Cas9 expression. Log-rank test. (B) Survival curves of primary (n = 9) and secondary (n = 17) recipient mice transplanted with double sgKmt2d/sgTrp53-targeted HSPCs with constitutive Cas9 expression. Log-rank test. (C) Survival curves of recipient mice transplanted with mock (n = 6), sgKmt2d (n = 6), sgTrp53 (n = 6), and double-targeted (n = 6) HSPCs with GC B-cell–specific Cas9 expression. Log-rank test. (D) Volcano plot showing differentially expressed genes between single sgKmt2d-targeted (n = 7) and double sgKmt2d-targeted/Trp53-altered (n = 17) B-cell lymphoma. Genes with q value of <0.2 and |log2(fold change)| of >2 are considered significant and colored red (gained) or blue (lost). (E) Yap1 mRNA expression measured by RNA-seq in samples with (n = 17) or without (n = 7) Trp53 alterations in sgKmt2d-targeted B-cell lymphoma. Brunner-Munzel test. (F) YAP1 and pYAP1 protein expression measured by capillary electrophoresis-based immunoassay in representative samples with or without Trp53 alterations in sgKmt2d-targeted B-cell lymphoma. Lamin B1 is used as a loading control. Representative of 3 independent experiments. ∗P < .05; ∗∗P < .005; ∗∗∗P < .0005. mRNA, messenger RNA; pYAP1, phosphorylated YAP1; TPM, transcripts per million.
