Fig. 1.

Isolation and differentiation of c-Kit+/Sca-1- cells. a: Morphology of differentiated ESC (ai) and flow cytometry strategy for the isolation of c-Kit+/Sca-1- subpopulation (red box, aii). Addition of VEGF and application of shear induced endothelial phenotype (EC, aiii) while PDGF induced smooth muscle cells phenotype (SMC, aiv). b: VEGF treated cells overexpressed EC markers as shown by real time-PCR (bi), immunocytochemistry (bii) and flow cytometry (biii). Functional differentiation was assessed by in vitro tube formation assay (biv). c: PDGF treatment induced expression of SMC markers, as shown at mRNA level (ci) and by immunocytochemistry (cii) and western blot analysis (ciii). Data are expressed as fold-change over Day 0 (dotted lines). Scale bars: 200 μm (a), 25 μm (bii), 100 μm (biv), 50 μm (cii) *P < 0.05; **P < 0.01; ***P < 0.001 vs. Day 0. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)