Figure 3: EndoMT is a key process in aortic valve development and is impaired by GATA4 deficiency.

(a) Western blot of GATA4 and GAPDH from control and GATA4 sgRNA ECs. GATA4 sgRNA ECs were differentiated from iPSCs transfected with px458 with GATA4 sgRNA and enriched by GFP. Control ECs were derived from iPSCs with px458 and enriched by GFP. An uncropped version is presented in Supplementary Fig. 9. Lower panel: quantification of western blot data. The data were normalized to control ECs. Experiments were repeated three times; averages and standard derivations were plotted. (b) Western blot of SMA and GAPDH from control ECs, control ECs undergoing EndoMT, GATA4 sgRNA ECs and GATA4 sgRNA ECs undergoing EndoMT. An uncropped version is presented in Supplementary Fig. 10. Lower panel: quantification of western blot data. The data were normalized to control ECs undergoing EndoMT. Experiments were repeated three times; averages and standard derivations were plotted. (c) Numbers of mesenchymal cells from control and GATA4 sgRNA in collagen gel assay. The data were normalized to control. Experiments were repeated three times; averages and standard derivations were plotted. (d) Immunofluorescence staining of SMA and CD31 of the control and GATA4 sgRNA undergoing EndoMT. Scale bars, 50 μm. EC, endothelial cell; EndoMT, endothelial-to-mesenchymal transition; iPSC, induced pluripotent stem cell; kDa, kilodalton; MW, molecular weight. *P<0.05; **P<0.01.