Figure 2
T63 increases osteoblast differentiation in MC3T3-E1 and C3H10T1/2 cells. (a) ALPL activity. MC3T3-E1 (left panel) or C3H10T1/2 (right panel) cells were cultured in OS medium, and treated with different concentrations of T63 for the indicated time before subjected to ALPL assay. *p < 0.05, **p < 0.01, ***p < 0.001 versus respective control (n = 4). (b) T63 increased Alpl mRNA expression. The cells were treated with T63 for 12 days and Alpl mRNA was analyzed by semi-quantitative PCR. Full-length gels are shown in Supplementary Fig. S6a. (c) Effect of T63 on the mRNA expression of Bglap, Spp1, Runx2 and Bmp2. The cells were treated with T63 (5 μM) for 12 days before qRT-PCR. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (n = 3). (d) Osteoblast mineralization. The cells were cultured in OS medium and treated with T63 for 21 days before Alizarin Red S staining (Scale bar: 50 μm). The quantitation of calcified nodules were shown as means ± SD, **p < 0.01, ***p < 0.001 versus control (n = 3).
