Fig. 2

Hematopoietic differentiation of the H1-GATA2 ^−/− ES cell line. a CFUs of H1 or H1-GATA2 ^−/− derived CD34⁺ cells. H1 or H1-GATA2 ^−/− cells were co-cultured with OP9 cells for 9 days. The CD34⁺ HPCs were isolated by FACS for CFU generation. Error bars represent mean + SEM of the mean of samples from nine independent experiments. b-d Time course analysis of blood differentiation of H1 and H1-GATA2 ^−/− cells upon co-culturing with OP9. The expression of surface markers CD34, CD43, and CD31 on H1 or H1-GATA2 ^−/− cells co-cultured with OP9 for the indicated time was analyzed by FACS (left and middle panels). The right panels are box plots from ten independent experiments on the percentage of indicated populations on day 8 of OP9 co-culturing. Asterisks indicate statistical significance determined by t test: ***p < 0.001. e Time course analysis of the expression of indicated genes of H1 and H1-GATA2 ^−/− cells upon co-culturing with OP9. The gene expression was analyzed by qRT-PCR by using GAPDH as an internal reference. f HPCs with CD34⁺CD31⁺CD43⁺ were developed from CD34⁺CD31⁺CD43⁻ HEs. CD34⁺CD31⁺CD43⁻ populations were sorted out at day 8 of differentiation and replated on OP9 for one additional day and analyzed by FACS. g FACS analysis of CD114 and KDR on HEs from H1 or H1-GATA2 ^−/−. h H1 or H1-GATA2 ^−/− derived HEs produced endothelial cells. Left: Morphology of endothelial cells; middle: immunostaining of CD31 on endothelial cells; right: capillary structure formation by endothelial cells