Sorting stage-matched F and Aerythroblasts. (A) Sorting scheme for HUDEP2 and CD34⁺ hematopoietic stem and progenitor cell (HSPC)-derived cells. Cells are gated using a viability marker and then matched for forward scatter, CD71, and CD235 expression. After sorting, cells generally have ∼95% to 99% purity. (B) RT-PCR validation of HUDEP2 F and A cell sorts from 3 HUDEP2C clones. Data are normalized to the average of 3 housekeeping genes and shown relative to A cells. (C) Western blot validation with sorted cells from 3 HUDEP2 clones. (D) ddRT-PCR analysis of globin transcripts in sorted F and A cells, normalized to AHSP transcripts. Percentage of HBG transcripts (HBG/[HBG + HBD + HBB] × 100) is also shown. All data are shown as individual points, mean, and standard deviation (SD) for 3 samples. ***P < .001; **P < .01. NS, not significant.