RBM12 depletion in primary human CD34⁺HSPCs does not impair erythroid maturation. CRISPR/Cas9 depletion of RBM12 in human CD34⁺ HSPCs via 2 independent sgRNAs (n = 3 biological replicates). (B, C, E, and F) Plotted are means ± standard deviation. (A) Western blot analysis with indicated antibodies at day 15 of differentiation. (B) Growth curve displaying total cell number from day 8 to 12 of differentiation. (C) Left: Representative contour plots of CD71 vs CD235ab at day 15 of differentiation. Right: Quantification of CD71_loCD235ab⁺ and CD71_loCD235ab⁺/HbF⁺ cells. ∗P < .05 by Student’s t test. (D) Representative May-Grünwald Giemsa images of HSPCs at day 15 of differentiation. (E) qRT-PCR analysis of globin genes at day 12 of differentiation. Results are normalized to HPRT1. ∗P < .05 by Student’s t test. (F) qRT-PCR analysis of differentiation markers GATA1, BAND3, and ALAS2 at day 12 of differentiation. Results are normalized to HPRT1.