Fig. 3

TNFα inhibits osteogenesis of rMSCs. a–c rMSCs were treated with TNFα (1 and 10 ng/ml) in OIM for 5 days, then gene expressions were detected by real-time PCR. β-actin was used as loading control. The experiments were repeated three times. **p < 0.01 compared with OIM. d Levels of Runx2 were analyzed by western blot with indicated antibodies. e Quantification of the band intensity using ImageJ software. Data are presented as mean ± SD (n = 3, *p < 0.05). f ALP in rMSCs was stained with BCIP/NBT kit after the cells were incubated with different concentrations of TNFα in OIM for 7 days. g The ALP activity of rMSCs was measured after they were incubated with different dosages of TNFα in OIM for 7 days. *p < 0.05 compared with Con (α-MEM), ^#p < 0.05, ^##p < 0.01 compared with OIM. h rMSCs were treated with TNFα (1 and 10 ng/ml) in OIM for 14 days, then the mineralized nodules were stained by Alizarin Red S. i The mineralization was quantified by extraction of Alizarin Red S dye with 10% CPC. **p < 0.01 compared with Con, ^##p < 0.01 compared with OIM