FA gene complementation rescues hematopoietic deficits. (A) FANCD2 Western blot on lysates from FANCD2i hESCs infected with pMMP-puro hFANCD2 or pMMP-puro (empty vector). (B) FANCAi hESCs infected with pMMP-puro hFANCA or pMMP-puro were treated with 2mM HU for 24 hours and lysates were subjected to Western blotting for FANCD2. FANCD2-L is the active, monoubiquitinated form of FANCD2-S. (C-D) FACS analysis of day 20 EB-derived cells for the percentage of CD45⁺ cells. Percentage CD45 is normalized to FANCD2i + hFANCD2 (C) or FANCAi + hFANCAi (D) and presented as ± SEM. n = 4, 6. (E-F) Total hematopoietic CFUs from day 20 EB-derived cells plated in H4434 media (StemCell Technologies) scored at day 14 after plating. Total CFUs are normalized to FANCD2i + hFANCD2 (E) or FANCAi + hFANCAi (F) and presented as ± SEM. n = 4, 6. (G) Venn diagram showing HOX genes with decreased expression in day 20 EBs. Eighty-four HOX genes were analyzed by quantitative PCR array (SABiosciences) and the top 8 genes, all with fold reduction greater than −1.5 relative to gene complement control, were included. FANCD2i + empty vector was compared with FANCD2I + hFANCD2 and FANCAi + empty vector was compared with FANCAi + hFANCA to generate the list of genes. Data are provided in supplemental Table 3. ***P < .01; *.03 < P < .05.