KDM5A KO in APL cell line NB4 prohibited proliferation, colony formation, and induced differentiation. (A) Proliferation assay of NB4 cells transduced with 2 control sgRNAs (Ctrl) and 2 KDM5A sgRNAs (KDM5A KO). Cell counts were measured at the indicated days after seeding. Data are shown as means ± standard error of the mean of triplicate analysis. The counts of KDM5A-KO cell lines at day 4 were compared with those in control cell lines. Statistical significance was determined by two-way analysis of variance. ****P < .0001. The P values of both KDM5A-KO groups were < .0001. (B-C) Bar graph and photos showing colony formation of the indicated NB4 cell lines using a 14-day methylcellulose assay. Colony counts were compared between 2 control cell lines and 2 KDM5A-KO cell lines, and statistical significance was determined by two-way analysis of variance. ***P < .001. (D) RT-qPCR analysis of differentiation markers CD11b, CD11c, and CD14 in the indicated cell lines. (E-F) Differentiation marker CD11b level in NB4 cells transduced with control sgRNAs and KDM5A sgRNAs measured by using flow cytometry. The statistical significance of RT-qPCR and percentage of CD11b⁺ cells was determined by two-way analysis of variance. ***P < .001. FACS, fluorescence-activated cell sorting.