Fig. 2.

Effects of FOXO3 on BM-MSC osteogenic differentiation factors were detected. a, b ALP levels in BM-MSCs after up-regulating or down-regulating FOXO3 were quantified with ELISA. c Effects of up-regulating or down-regulating FOXO3 on BM-MSC osteogenic differentiation were detected with Alizarin Red Staining, under 100 × magnification. d–g Relative protein and mRNA expressions of factors related to osteogenic differentiation (RUNX2; OCN; Smad4) were measured with Western blot and qRT-PCR. GAPDH was an internal control. All the experiments have been performed in independent triplicate and the experimental data were expressed as mean ± standard deviation (SD). ⁺⁺P < 0.01, ⁺⁺⁺P < 0.001, vs. NC; ^{^^^}P < 0.001, vs. siNC. RUNX2 Runt-related transcription factor 2, OCN Osteocalcin, Smad4 SMAD family member 4, siNC small interfering RNA for negative control