CDK6 acts as a central node in apoptotic network, NF-κB signaling, and HSC activation in JAK2^V617Fmutant LSKs. (A) Heat map shows differentially expressed genes from LSKs of indicated genotype (adjusted P ≤ .05; absolute log2-fold changes >1; contrast: VavCre; JAK2^V617F; Cdk6^−/− vs VavCre; Jak2^+/+; Cdk6^+/+). Regularized log-transformation of count data was used as input for the heat map. For full data set, see supplemental Figure 9B and supplemental Table 1. Color code: red, upregulation; blue, downregulation. n = 3 per genotype. (B) Gene set enrichment analysis for gene expression signatures of apoptosis and TNF-α signaling via NF-κB in LSKs of VavCre; JAK2^V617F; Cdk6^−/− compared with VavCre; JAK2^V617F; Cdk6^+/+. For full data set, see supplemental Figure 8D. Color code: red, upregulation; blue, downregulation. FDR, false discovery rate; NES, normalized enrichment score. (C-D) Expression of indicated genes was determined by quantitative real time polymerase chain reaction in LSKs of respective genotype. Relative expression levels were normalized to the housekeeping genes Rplp0 and Hprt. Results are presented as means + SD (n = 3 per genotype). One-way ANOVA with subsequent Bonferroni posttest was used, and significance is indicated. *P < .05; **P < .01; ***P < .001; ****P < .0001. (E) Levels of indicated proinflammatory cytokines were measured in plasma samples of respective genotype. Error bars indicate + SD (n ≥ 5 per genotype). One-way ANOVA with subsequent Bonferroni posttest was used, and significance is indicated *P < .05. IFN, interferon; N/A, not detected. (F) Bubble plot showing relative mRNA levels for indicated genes determined by quantitative real time polymerase chain reaction in LSKs (n = 3 per genotype). Relative expression levels were normalized to the housekeeping genes Rplp0 and Hprt. Circled area corresponds to relative expression. Significance is indicated in supplemental Figure 9C.