Fig. 3

Characterization of the occurrence of the EHT during the hematopoietic specification. A Morphological features of adherent cells and suspension cells climbed out from aforementioned adherent cells at the 11th day after the hematopoietic differentiation from hESCs (10× magnification, scale bars 100 μm, far left one photo; 20× magnification, scale bars 50 μm, right three photographs). B CLSM images of the immunostaining for VE-Cadherin and CD45 in the differentiated cells at day 11 undergoing the EHT. The colors of green, red and blue represent the staining for CD45, VE-Cadherin and nuclei, respectively. (Original magnification, 20×, scale bar 40 μm, upper panel; 40×, scale bar 50 μm, lower panel.). C Flow analysis for HEPs (CD31⁺CD45⁺ cells or CD144⁺CD45⁺ cells) at the 11th day during the hematopoiesis. D The dynamic gene expression of the indicated HSC-specific transcription factors by RT-qPCR during the hematopoiesis. E The transduced efficiency of lentivirus containing EGFP and mCherry reporters in hESCs was evaluated by fluorescent microscopy. (Original magnification, 10×, scale bar 100 μm). F The aggregate-forming ability of engineered hESCs seeded on the low adhesion plate was not affected by the transduction with the lentivirus. (Original magnification, 4×, scale bar 10 μm) G The generation of EGFP and mCherry double-positive hematopoietic cells during the development of the hematopoiesis was visualized by living cell fluorescence trace system. (Original magnification, 4×, scale bar 100 μm). Data represent the mean ± SEM. *p < 0.05 and ns (no significance)