Bahcc1 depletion prolongs development of MLL-ENL-induced leukemia. (A) Experimental strategy using secondary BMT. Primary leukemic cells expressing MLL-ENL together with EGFP were harvested, followed by retroviral transduction with shRNA/Kusabira-Orange (KO) expressors. Leukemic cells highly expressing KO were sorted and immediately transplanted into secondary recipient mice. (B) Relative-relative expression levels of Bahcc1 in shRNA-transduced leukemic cells. shLuc, shRNA against luciferase; shB-2, shRNA against Bahcc1. (C) Survival curves of mice transplanted with Bahcc1-depleted (shB-2; n = 12) or control (shLuc; n = 10) leukemic cells. Data from 3 independent experiments were combined. (D) Spleen weights of moribund mice. (E) Representative FACS plots of BM cells from moribund mice. GFP/KO doubly positive cells were gated as shown in the top panels, followed by subsequent analyses for c-kit, Gr-1, and CD11b expression. (F) Relative-relative expression levels of Bahcc1 by RT-qPCR in BM cells from moribund mice. Bar graphs show the mean with SD of at least 3 independent experiments. ns, not significant (log-rank test for panel C and unpaired Welch t tests for panels D-E). Wt, wild-type.