Fig. 5

FoxC1 initiates Oct4 activation. A, B FoxC1 overexpression in ECs increases the expression of Oct4 mRNA and protein in cocultured MSCs. The indicated core stemness factors were analyzed in the MSCs cocultured with adFoxC1-treated ECs, Ctrl-ECs, or siFoxC1-treated ECs. C, E The mRNA and protein expressions of Oct4-relative pro-angiogenic factors Ang1, bFGF and VEGF, anti-apoptic factor Bcl-2, and pro-apoptic factors Bax and caspase3 were detected by immunoblotting (C) and real-time RT-PCR (E) in these MSCs cocultured with adFoxC1-treated ECs, Ctrl-ECs, or siFoxC1-treated ECs. D Correlation analysis of Oct4 positive MSCs cocultured with adFoxC1-treated ECs and FoxC1 expression in the ECs transfected with adFoxC1 were analyzed by quantitative immunofluorescence and western blotting. F Representative immunostaining in MSCs cocultured with adFoxC1-treated ECs or siFoxC1-treated ECs showed Oct4 expression was consistent with VEGF and Bcl-2 expression, but opposite to caspase 3 expression. The nuclei of MSCs were stained blue using DAPI. Scale bars = 50 µm. All data are the means ± SEM. p < 0.05: *versus adFoxC1 group, ^†versus Ctrl group (n = 10 per group). Welch ANOVA analyses were performed in A and E