YTHDC1 KD inhibits leukemic potentials of human primary AML cells. Growth curve of primary AML cells from 4 different patients (A-D) and CD34⁺ cells (E) from healthy donors expressed Scramble shRNA (Scr), YTHDC1 shRNA1# (Ysh1#), and YTHDC1 shRNA2# (Ysh2#). CD34⁺ cells are mixed cells from 3 healthy donors. The experiments were performed in triplicate. The P value was detected by 1-way analysis of variance followed by multiple comparisons vs the Scr group at day 10 (for patient cells) or day 7 (for CD34⁺ cells). (F) Flow cytometric analysis of apoptosis frequency of CD34⁺ cells in patients with AML and CD34⁺ cells from healthy donor expressed Scramble shRNA (Scr), YTHDC1 shRNA1# (Ysh1#), and YTHDC1 shRNA2# (Ysh2#). (G) Colony-forming units of AML patient cells and CD34⁺ cells from healthy donor expressed Scramble shRNA (Scr), YTHDC1 shRNA1# (Ysh1#), and YTHDC1 shRNA2# (Ysh2#). A total of 50 000 cells per well were used for patient cell input and 2500 cells per well for healthy donor CD34⁺ cell input. Flow cytometric analysis of differentiated cell frequency of AML patient cells (H-J) and CD34⁺ (L) cells from healthy donor expressed Scramble shRNA (Scr), YTHDC1 shRNA1# (Ysh1#), and YTHDC1 shRNA2# (Ysh2#). (K) Gating strategy. (M) Wright-Giemsa–stained AML cell morphology, with differentiated cells indicated by red arrows; bar = 20 μM. (N) Flow cytometric analysis of the percentage of human leukemia cells (CD45⁺) in NSG mice–transplanted MOLM-13 cells expressed Scramble shRNA (Scr), YTHDC1 shRNA1# (Ysh1#), and YTHDC1 shRNA2# (Ysh2#). (O) Kaplan-Meier survival analysis of NSG mice–transplanted MOLM-13 cells expressed Scramble shRNA (Scr), YTHDC1 shRNA1# (Ysh1#), and YTHDC1 shRNA2# (Ysh2#). Data are presented as mean ± standard deviation; Student t test or log-rank (Mantel-Cox) test for survival curve. Experiments were performed in triplicate. *P < .05; **P < .01; ***P < .001.