Fig. 3

MSX2 deletion augments hematopoietic differentiation of hESCs by facilitating EHT. a Flow cytometry analysis of the percentage of APLNR⁺ mesoderm cells from H1 WT and H1 MSX2^−/− cells at day 2 of hematopoietic differentiation. b Flow cytometry analysis of the percentage of CD31⁺CD34⁺ HEPs from H1 WT and H1 MSX2^−/− cells at day 5 of hematopoietic differentiation. c Flow cytometry analysis showing the percentage of CD31⁺ cells (left) and CD43⁺ subpopulation gated on CD31⁺ cells (right) from H1 WT and H1 MSX2^−/− cells at day 8 of hematopoietic differentiation. d Schematic overview showing the experimental design to determine the hematopoietic potential of CD31⁺CD34⁺ HEPs. HEPs were sorted at day 5 of hematopoietic differentiation and seeded into the hematopoietic culture for 3 days before immunofluorescence and flow cytometry analysis. e Representative photomicrographs of cobblestone-like cells generated from H1 WT and H1 MSX2^−/− HEPs. f Representative immunofluorescence images of CD43⁺ HPCs (red) generated from H1 WT and H1 MSX2^−/− HEPs. Nuclei were stained with DAPI (blue). g Representative flow cytometry dot plots (left) and statistical analysis (right) showing the generation of CD43⁺ HPCs emerging from H1 WT and H1 MSX2^−/− HEPs. h Flow cytometry analysis showing the percentage of CD43⁺ subpopulation gated on CD31⁺ cells from H1 WT and H1 MSX2^−/− cells with or without MSX2 overexpression at day 8 of hematopoietic differentiation. Results are shown as means ± SD (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001