Figure 3

RTKIs reduce CPC potential to differentiate. (a) Real-time qPCR analysis of CPC early differentiation markers: Nkx2.5 reduced by 10 µM ST for 24 h and (b) but without changes to Nkx2.5 protein expression in CPCs post ST treatment. (c) Real-time qPCR analysis of CPC early differentiation: PECAM-1 reduced by 5 µM ST and Nkx2.5 reduced by 5 µM IM, 0.5 µM SM or 5 µM ST at 7 days; (d) protein expression was also assessed. Directed differentiation of CPCs in vitro determined that: (e) percentage of CPCs expressing α-sarcomeric actin was reduced by ST (n = 4), (f) representative images (scale bar = 100 µm); (g) percentage of CPCs expressing smooth muscle actin was reduced by SM (n = 4), (h) representative images (scale bar = 100 µm); (i) percentage of CPCs expressing von Willebrand factor was reduced by SM (n = 4), (j) representative images (scale bar = 100 µm). All data are mean ± SEM; significance was determined by ANOVA with Holm-Šídák post hoc test to identify differences (*p < 0.05; **p < 0.01; ***p < 0.001). Blots cropped to relevant data, with optical density normalized to the loading control protein; full blot images presented in Supplementary Fig. 6.