GPS2 expression is upregulated during erythroid differentiation of CD34⁺cells. (A) Flow cytometry gating strategy for HSPCs in CD34⁺ cells isolated from human UCB: HSCs (CD34⁺CD38⁻), CMPs (CD34⁺CD38⁺CD123⁺CD45RA⁻), GMPs (CD34⁺CD38⁺CD123⁺CD45RA⁺), and MEPs (CD34⁺CD38⁻CD123⁻CD45RA⁻). (B) The relative expression of GPS2 and GATA1 in HSCs, CMPs, GMPs, and MEPs isolated from human UCB was analyzed by real-time PCR (normalized to GAPDH levels). (C) Representative microscopy images of BFU-Es and CFU-GMs cultured from human UCB CD34⁺ cells. (D) Real-time PCR analysis of GPS2 and GATA1 expression in BFU-Es and CFU-GMs (normalized to GAPDH levels). (E-G) Human UCB CD34⁺ cells were isolated and cultured for 2 days in expansion medium, which was then changed to erythroid differentiation medium for the indicated times. Flow cytometry plots showing the expression of CD71 and glycophorin A (GPA) (E). Real-time PCR (F) and western blot (G) analysis of the expression of GPS2. All values are mean ± SEM (n = 3 replicates). *P < .05, ***P < .001; 2-tailed unpaired t test.