Fig. 2: One step differentiation of CPCs to VELs. | Communications Biology

Fig. 2: One step differentiation of CPCs to VELs.

From: Generation and characterization of cardiac valve endothelial-like cells from human pluripotent stem cells

Fig. 2

a Schematic of differentiation hPSC-derived CPCs to ECC and VEC-like cells. b The qRT-PCR analysis of the selected ECC genes for day 6 hPSC-derived cells. Note that the combined treatment with VEGF, BMP4, and TGFb1 leads to a marked upregulation of ECC genes. c Time-course gene expression analysis of the indicated genes during hPSCs differentiation to CPCs and CPCs to VEC-like cells. The ECC-enriched genes are highlighted by a red dashed box, and the VEC-enriched genes are highlighted by a blue box. d IF staining of day 6 hPSC-derived cells, showing double positive cells for NOTCH4/VE-cad, DLL4/VE-cad and CD31/JAG1, respectively. Scale bar: 25 μm. e Percentage of NOTCH4/VE-cad-, DLL4/VE-cad- and CD31/JAG1-double-positive cells of (d), quantified by ImageJ. Bar graph represents double positive cells ± S.D of three independent experiments. f Flow cytometry results showing the percentage of JAG1/VE-cad double positive cells. Left: epitope controls; Right: JAG1/VE-cad antibodies. g IF staining of day 8 hPSC-derived VELs, showing double positive cells for HEY2/VE-cad, PROX1/VE-cad and TBX2/VE-cad, respectively. Scale bar: 25 μm. h WB analysis for hPSC-derived CPCs and day 8 hPSC-derived VELs, with the indicated antibodies. i Flow cytometry results showing the percentage of NFATc1/VE-cad and TBX2/VE-cad double positive cells. The paired t test in Graphpad software was used for the statistical analysis. Significant levels are: *p < 0.05; **P < 0.01; ***P < 0.001. Shown are representative data in (d, g).

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