Figure 4

TUG-891, an agonist of GPR120, significantly promotes the osteogenesis of BMMSCs at high concentrations.

BMMSCs were incubated for 14 days of osteogenesis in the presence of different concentrations of TUG-891 [0 (Con), 0.1, 0.5, 1, 5, 10, 30, 50, 100 μM]. (A) ALP (scale bar = 200 μm) and (B) Alizarin red (scale bar = 80 μm) staining of cells. (C) Effect of TUG-891 on the ALP activity of the osteogenic differentiation of BMMSCs. The control value for ALP activity was 0.432 ± 0.015 units/mg protein. (D) Effect of TUG-891 on the mineralization of the osteogenic differentiation of BMMSCs. The control value for mineralization was 0.920 ± 0.016 OD. (E) Effect of TUG-891 on the osteogenic mRNA expression of Alp, Runx2 and Ocn. (F) Western blot analysis of ERK1/2, pErk1/2, Ras, Ras-GRF, p38 and pp38. The expression of each target gene was calculated as the relative expression to β-actin and is represented as normalized fold expression. Data are represented as the mean ± SD of 3 independent experiments. *P < 0.05 and **P < 0.01 compared with the control group.