FANCD2i hESCs demonstrate significant reductions in hematopoietic CFUs. (A) Representative light and fluorescence micrographs of hematopoietic colonies. BFU-E indicates blast-forming unit erythroid; CFU-GM, colony-forming unit granulocyte/monocyte; and CFU-M, colony-forming unit macrophage. (B-C) Total hematopoietic CFUs from day 20 EB-derived cells plated in H4434 media (StemCell Technologies) without (B) or with (C) 5nM MMC and scored at day 14 after plating. Total CFUs are normalized directly to LUCi (B) or first normalized to baseline untreated total CFUs to assess MMC hypersensitivity of hematopoietic progenitors (C). Data are presented as mean ± SEM. n = 6, 3. (D) Normalized ratio of gamma-globin expression to epsilon-globin expression by qRT-PCR from CD45⁺ population of day 20 EB-derived cells. Data are presented as mean ± SEM. n = 6. (E-G) qRT-PCR analysis of gene expression from CD45⁺ population of day 20 EBs. Expression is relative to LUCi expression levels after normalization with GAPDH. Data are presented as mean ± SEM and n = 6. ***P < .01; **.01 < P < .03; *.03 < P < .05.