Figure 6: FOXO1 promotes proliferation and osteogenic differentiation through the miR-424/FGF2 pathway under oxidative stress.
hASCs were transfected with a miR-424 mimic and a miR-424 inhibitor for 24 h. A mimic NC (negative control) and an inhibitor NC were used as a negative control. Thereafter, the cells were incubated with basal medium (BM) or osteogenic medium (OM) and were continuously exposed to H2O2 until the indicated times. (a) Western blot showed the protein expression of FGF2 at 3 days after the knockdown of FOXO1 and the cotransfection of FOXO1 siRNA or NC-siRNA and miR-424 inhibitor or inhibitor NC. (b) The proliferation of hASCs was evaluated by a MTS assay after the knockdown of FOXO1 and the cotransfection of FOXO1 siRNA or NC-siRNA and a miR-424 inhibitor or inhibitor NC. (c,d) The expression of RUNX2 and ALP were evaluated after the knockdown of FOXO1 and the cotransfection of FOXO1 siRNA or NC-siRNA and a miR-424 inhibitor or inhibitor NC and then osteogenic induction for 7 days. H2O2 + OM was used as the control group. *P < 0.05, **P < 0.01, #P < 0.05, ##P < 0.01 compared to the si-FOXO1 + inhibitor NC group, ns, not significant. The results indicate the mean ± SD of triplicate experiments. The images shown are representative of three independent experiments. (c) A schematic representation of the main conclusions of this study.
