Cell cycle and apoptosis analysis in FANCA-expressing and -deficient IPSC-derived hematopoietic cells. (A-B) HPCs were isolated after the completion of EHT culture, at which time they were isolated and stained with DAPI and Ki67 to quantify the cell cycle. The percentage of cells in each phase of the cell cycle was quantified (n = 5 biologic replicates; each phase analyzed by a paired Student t test, results are presented as mean ± SEM). (C) Expression level of the CDKN1A transcript was measured by quantitative PCR and expression normalized to β-actin mRNA. Expression was compared by Student t test with 6 biologic replicates. (D) GSEA analysis of RNA sequencing data. (E-F) Subcellular localization of p53 was analyzed by immunofluorescence staining and nuclear fluorescence signal quantified normalized to DAPI signal (scale bar, 10 μm; results aggregated from 3 biologic replicates, compared by a Student t test, results presented as mean ± SEM; n = 240 Dox cells and 111 No dox cells quantified). (G) p53 protein was measured in day 8 EHT HPCs from 2 independent cell lines under the indicated conditions.