Fig. 7.

FOXO3 reversed the effects of miR-223-3p on BM-MSC osteogenic differentiation. a, b ALP levels in BM-MSCs after overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p were quantified with ELISA. c, d Effects of overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p on BM-MSC osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. e–h Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) after overexpressed FOXO3 or siFOXO3 and up-regulated or down-regulated miR-223-3p were measured with Western blot and qRT-PCR. GAPDH was an internal control. All experiments have been performed in triplicate and experimental data were expressed as mean ± standard deviation (SD). ⁺P < 0.05, ⁺⁺P < 0.01, ⁺⁺⁺P < 0.001, vs. NC; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, vs. FOXO3; ^▲▲▲P < 0.001, vs. 3-MA; ^{^}P < 0.05, ^{^^}P < 0.01, ^{^^^}P < 0.001, vs. siNC; ^&&P < 0.01, ^&&&P < 0.001, vs. siFOXO3; ***P < 0.001, vs. RAPA