sCFU-E cells exhibit higher growth potential and express lower levels of erythroid-committed genes compared with CFU-E cells. (A) Histologic staining of sorted BFU-E, sCFU-E, and CFU-E cells. (B) Quantification of forward scatter (FSC) median fluorescence intensity, an indicator of cell size, by flow cytometry. (C) sCFU-E cells generate unifocal colonies on day 2. Colonies generated from sCFU-E cells are larger than those from CFU-E cells. (D) Quantification of area per colony and average cell number per colony in (C). (E) Sorted sCFU-E cells generate more progenies than CFU-E cells in vitro. (F) Comparison of sCFU-E and CFU-E cell transcriptome by RNA-seq. The number of genes with expression greater (up) or less (down) than 1.5-fold are indicated. (G) Relative expression of indicated genes in sCFU-E vs CFU-E cells from RNA-seq data. (H) Expression of indicated genes in sorted BFU-E, sCFU-E, and CFU-E by qPCR. Gene expression is normalized first to β-actin and then to expression in BFU-E cells. Significant differences between sCFU-E and CFU-E cells are specified. Data represent mean ± SD. Statistically significant differences indicated on top of each bar are in comparison with BFU-E cells. ∗P < .05; ∗∗P < .01, Student’s t test or 1-way ANOVA.