DPY30 KD promoted erythroid maturation of human HPCs and erythroleukemia cells. Human CD34⁺ cells were depleted of DPY30 by 2 different shRNAs (#2 and #5), and were induced to differentiate into erythroid lineage in liquid culture with appropriate cytokine conditions (containing 1 ug/mL EPO). (A) Indicated cell surface markers were analyzed and quantified by flow cytometry at different days during differentiation. Averages ± SD from 4 independent biological repeats are plotted. *P < .05; ***P < .001 (Student t test). (B) Image of pelleted cells on day 14 of erythroid differentiation. (C) The expression of hemoglobin genes before the differentiation assays was determined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Averages ± SD from 4 biological repeats are plotted. *P < .05; **P < .01 (Student t test). (D-E) Ten days after DPY30 KD (by sh#2 viral infection), K562 cells were checked by western blot analysis for KD and effect on H3K4 methylation (D), and stained by benzidine (E, top) and pelleted for image (E, bottom). (F) Expression of HBA1, HBG1, and DPY30 in control and DPY30-KD K562 cells was monitored by qRT-PCR against ACTB. Completion of selection, indicated by loss of viability of most uninfected cells after culturing in the presence of puromycin, was 4 days after viral infection. Averages ± SD from 3 independent biological repeats are plotted. *P < .05; **P < .01 (Student t test). (G) Growth of control and DPY30-KD K562 cells was monitored daily. Averages ± SD from 3 biological repeats are plotted.