Fig. 1

Identification of mesodermal specification during the hematopoietic differentiation from hESCs. A Schematic representation for the generation of definitive HPCs from hESCs. B The expression of mesoderm surface marker (KDR) and corresponding isotype controls (shown in blue) during the in vitro hematopoietic development from hESCs. The lower left panel showed representative images of EBs at different time points. (Original magnification, 10×, Scale bar, 100 μm). C Generation of EpCAM⁻CD56⁺ mesoderm-enriched cells shown by flow cytometry at days 0, 2 and 4 employing hematopoietic differentiation protocol. D Relative expression levels of EpCAM and CD56 genes were determined and quantified by RT-qPCR in the bar graphs. E Expression analysis of pluripotent genes (SOX2 and OCT4) and mesodermal genes (CDX4, KDR, BMP4 and Brachyury) for different cell populations at different time points during the hematopoietic specification from hESCs by RT-qPCR. Data represent the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 and ns (no significance). F EBs were fixed with PFA and subjected to immunofluorescent staining for KDR and CDX4 at day 4 of differentiation from hESCs. (Original magnification, 20×, scale bar 40 μm, upper panel; 40×, scale bar 50 μm, lower panels.)