Figure 5
Mesenchymal-to-epithelial Transition (MET) participated in the dedifferentiation from SMCs to PR-SMCs. (a) Data collected from RNA-Seq showed the overall suppression of mesenchymal genes and activation of epithelial genes in PR-SMCs compared with SMCs. Color bar indicates gene expression in scale. (b) Real-time PCR results showed that during SMCs to PR-SMCs dedifferentiation, mesenchymal markers α-SMA, Fibronectin, SNAI1 were downregulated and epithelial markers E-Cadherin, Claudin-1, Mucin 1 were upregulated (*p < 0.05, **p < 0.01, ***p < 0.001, n = 3). Control group refers to the SMCs transfected with empty lentivirus vector and maintained under the same reprogramming conditions. (c) The upregulation of epithelial marker E-Cadherin and suppression of mesenchymal markers SNAI1, N-Cadherin were confirmed at protein level by western blot analysis. (d) Real-time PCR result indicated that knockdown E-Cadherin with shRNA at day 2 of reprogramming in concert with the downregulation of CD34 in PR-SMCs at the mRNA level (**p < 0.01, n = 3). (e) When subjecting PR-SMCs with or without E-Cadherin suppression to endothelial differentiation, E-Cadherin knockdown PR-SMCs showed impaired capacity of endothelial markers induction (***p < 0.001, n = 3). Western blots were cropped for clarity. Examples of uncropped blots are found in Supplementary Figure 15.
