Figure 7.

rWNT5A and rWNT3A Regulate VSMC Phenotypic Switching. Representative western blots evaluating the expression of markers of synthetic or contractile VSMCs. VSMCs were incubated in rWNT5A or rWNT3A supplemented medium (both 100 ng/mL) for 24 h. (A) rWNT3A increased expression of PDGF-receptor-beta, collagen I, and β-catenin. rWNT3A decreased markers of contractile phenotypic differentiation (calponin, smoothelin). rWNT5A did not significantly reduce contractile markers. rWNT5A decreased PDGF-receptor-beta and markers of synthetic phenotypic differentiation (collagen I). rWNT5A also decreased β-catenin expression. Experiments were performed in triplicate. (B) Representative western blots evaluating expression markers of synthetic or contractile VSMCs. VSMCs were incubated in rWNT5A or rWNT3A in combination at the indicated concentrations (0, 50, 100, 200 ng/mL for each WNT) for 24 h. Taken together, these results suggest that under different concentrations of rWNT5A and rWNT3A, VSMCs up-regulate or down-regulate expression of the assayed phenotypic markers. Data are representative of one experiment performed in duplicate. Western blot band intensities were quantified and are presented as bar graphs (mean ± SEM) as band intensity or ratio of control normalized (protein of interest)/Tubulin. * = p < 0.05; ** = p < 0.01; *** = p < 0.001 significant difference between groups.