Fig. 5

ERK and AKT inhibition decreased the enhancing effect of ETV2 overexpression on osteogenesis of hDPSCs. a, b The addition of 50 μM PD98059 (a) or 20 μM LY294002 (b) reduced the levels of phosphorylated ERK (a) or AKT (b) due to ETV2 overexpression. c, d Results of qRT–PCR (c) and western blot analysis (d) revealed that PD98059 and LY294002 partially decreased the upregulated mRNA and protein expression of osteogenic markers due to ETV2 overexpression. e Representative images of immunofluorescence staining of OSX at day 3 after osteogenic induction and corresponding quantification. Scale bar: 50 μm. f ALP staining after osteogenic induction for 7 and 14 days and relative quantification of ALP activity. Scale bar: 200 μm. g ARS staining at days 14 and 21 of osteogenic induction and corresponding quantitative analysis. Scale bar: 200 μm. * P < 0.05, compared with DOX (−-) group; # P < 0.05, compared with DOX (+) group; & P < 0.05, compared with DOX (+) + PD98059 group; $ P < 0.05, compared with DOX (+) + LY294002 group. Error bars, mean ± SD (n = 3)