JMJD1C depletion triggers apoptosis. (A) Relative percentage of annexin V–positive MLL-AF9 cells at 3, 4, and 6 days after transduction with nontargeting control (shScr) or shRNAs targeting Jmjd1c (sh867 and sh868). Average of 3 independent experiments; error bars indicate standard deviation. (B) Relative percentage of annexin V–positive SEM cells over a time course starting from IPTG induction at day 0. Average of 3 independent experiments, error bars indicate standard deviation. (C) Western blot showing JMJD1C, poly ADP ribose polymerase, and caspase 3 levels in control (pLKO Scr) and JMJD1C-depleted SEM cells (shJ1C#i60 and shJ1C#i61). Vinculin was used as a loading control. (D) Number of SEM cells over a time course of JMJD1C KD induction by IPTG. Fresh IPTG was added at days 0, 2, and 4. (E) Cell cycle analysis of JMJD1C-depleted SEM cells (sh#i60) compared with control cells (Scr). Error bars indicate standard deviation (n = 3 for days 0 to 2; n = 2 for day 3). (F) Flow cytometry analysis of c-Kit and Mac1 expression in MLL-AF9 cells with shScr or shJmjd1c. (G) Relative mRNA levels of the indicated genes in cells transduced with shScr or shJmjd1c. (H) Representative images of May-Grünwald-Giemsa–stained MLL-AF9 cells transduced with shScr or shJmjd1c.