Fig. 3: Knockdown of NHE1 suppressed the growth of kinase-mutated AML.

A Representative western blot analysis showing successful NHE1 knockdown by shRNA in MV4-11, MOLM-13, and Kasumi-1. B–D In vitro NHE1 knockdown B reduced pHi, C induced apoptosis, and D suppressed proliferation in MV4-11, MOLM-13, and Kasumi-1 compared to scrambled shRNA control (n = 3). E–G In vitro NHE1 knockdown E suppressed the growth, F reduced pHi, and G induced apoptosis (normalized to scrambled shRNA control) in primary AML samples carrying FLT3 or RAS mutation, but not in AML with wildtype FLT3 or RAS (n = 13). H NHE1-KD suppressed the human leukemia engraftment of MV4-11 and MOLM-13 in NSG mice, compared to scrambled shRNA control, at week 5 and week 3 post transplantation respectively (n = 6–9). I Survival analysis of NSG mice engrafted with MV4-11 and MOLM-13 carrying NHE1-KD showed longer survival compared to those with scrambled shRNA control (n = 7–11). J NHE1-KD reduced ex vivo pHi MV4-11 engrafting in NSG compared to scrambled shRNA control (n = 6–7). K NHE1-KD suppressed human leukemia engraftment of primary AML samples with FLT3 or RAS mutation in NSG mice, compared to scrambled shRNA control, at week 12–16 post transplantation (n = 6). L Representative western blot analysis of kinase activation upon therapeutic inhibition in MV4-11 (quizartinib: 10–100 nM; ibrutinib: 5–10 μM; BRD7389: 5–10 μM) (the number showing the ratio of phosphorylated kinase to the corresponding total kinase normalized by the vehicle control. UD undetected).