Derepression of Cdkn1c in immortalized LT-HSCs with Bahcc1 depletion. (A-B) RT-qPCR of Cdkn2a, Cdkn2b, Cdkn1c, and Bahcc1 in immortalized LT-HSCs transduced with shRNA/KO expressors. Cells sorted on the basis of high KO expression 72 hours after transduction (A), and colony-forming cells (B) derived from the shRNA-transduced cells shown in Figure 3C were analyzed. (C) ChIP-qPCR of H3K27me3 around exon 1 of Cdkn1c using FLAG-tagged-MLL-ENL-immortalized KSL cells. Primer sets (1-4) are shown (top). Hoxc8 and CD11b were used as controls. shLuc, shRNA against luciferase; shB-2 and shB-8, different shRNAs against Bahcc1. Bar graphs show the mean with SD of at least 3 independent experiments. ^∗P < .05; ^∗∗P < .01; ^∗∗∗P < .001; ^∗∗∗∗P < .0001; ns, not significant. ANOVA followed by Dunnett multiple comparisons for panels A-B and unpaired Welch t tests for panel C.