Analysis of cytokine-induced in vitro hematopoietic differentiation of disease model iPSCs. (A) A schematic protocol using a liquid medium to induce hematopoietic differentiation in iPSCs, as described previously.¹⁴  (B) The entire cellular composition from the indicated iPSCs, stained with the indicated antibodies, was analyzed by flow cytometry. (C-E) The experiments in duplicate culture were repeated twice with different sets of iPSC clones, and mean ± standard deviation (SD) are shown. The P values were calculated using 1-way ANOVA with Tukey’s multiple-comparisons test. (C) Quantification of the percentage of CD34⁺ hematopoietic progenitor cells on day 6. (D) Quantification of the percentages of CD34⁺, CD34⁺CD45⁺, and CD45⁺ cells on day 10. (E) Quantification of the percentage of CD45⁺ hematopoietic cells on day 18. (F) FANCD2 foci were scored as the percentage of cells with >5 foci per cell. Mean ± SD in quadruplicate cultures are shown. The number of foci was evaluated by an IN Cell Analyzer 2000 with ≥3000 cells for each sample. Representative images of cells are shown in supplemental Figure 8. ADH5^−/−ALDH2^−/− iPSCs were excluded from this analysis because of extensive cell death. n.s., not significant.