Fig. 3

The Effect of PNS on EPCs is Mediated by the Wnt/β-catenin Signaling Pathway. a The mRNA expression of Wnt3a, Wnt5a, LRP5, β-catenin and TCF4 was detected by qRT-PCR. b Immunofluorescent microscopy was used to elucidate the cellular localization of β-catenin after treated with PNS. c, d The expression of β-catenin was assessed at the mRNA (c) and protein (d) levels in control cells, NC cells, and cells transduced with three different shRNA constructs targeting β-catenin. **p < 0.01 v.s. CTRL. e-f Effect of PNS treatment on nuclear enrichment of Wnt/β-catenin Pathway transcription factors. EPCs were exposed to increasing concentrations of PNS and incubated for 72 h. At the end of the incubation period, cytosolic and nuclear protein extracts were prepared and subjected to WB analysis. Protein levels of β-catenin were measured in nuclear and cytosolic protein fractions. * p < 0.01, ** p < 0.01 v.s. CTRL. g-h PNS promotes angiogenesis-related protein expression and Wnt/β-catenin signaling pathway activation. Western blot to evaluate the protein levels of VEGF-A, bFGF, VE-cadherin, DVL, GSK-3β and P-GSK-3β in indicated groups. The average levels were determined by ImageJ for three independent experiments. * p < 0.05, ** p < 0.01, ## p < 0.01 v.s. shRNA-β-catenin group