Expression profiling on Phf6-deficient HSCs. (A) Volcano plot showing gene-expression changes upon the Phf6 deletion. Red circles indicate genes that showed significant expression changes (q value < 0.05) in Phf6^Δ/y HSCs. (B) Differentially expressed genes and their fold-expression changes are shown as a heatmap. “α” shows genes included in the gene set “HALLMARK_TNFA_SIGNALING_VIA_NFKB.” (C) RT-qPCR analysis of the indicated genes in CD150⁺CD34⁻LSK and CD150⁺CD34⁺LSK cells. Hprt1 was used to normalize the amount of input RNA. Data are shown as the mean plus or minus SEM (n = 3). Significance is shown relative to VC. *P < .05, **P < .01, ***P < .001 by the Student t test. (D) GSEA performed using RNA-sequence data. GSEA plots of representative data are shown. Normalized enrichment scores (NES), nominal P values (NOM), and false discovery rates (FDR) are indicated. (E) Effects of TNFα on HSC proliferation. Freshly sorted HSCs were cultured under HSC culture conditions (10 ng/mL SCF and TPO) with various dosages of TNFα for 10 days. Data are shown as the mean plus or minus SEM. *P < .05; **P < .005; ***P < .0005 by the Student t test. TGF, transforming growth factor.