Figure 6.

ED-71 regulated MC3T3-E1 cells through Notch and Wnt/GSK-3β/β-catenin pathway. (A). The representative immunofluorescence staining images of β-catenin in CON, OM, DEX, DEX+ED-71 groups at 7 days. Bar, 200μm. (B). Notch1 immunofluorescence staining in CON, OM, DEX, DEX+ED-71 groups at 7 days. Bar, 200μm. (C). The mRNA levels of β-catenin and Notch1 detected by RT-qPCR after 7 days of culture. GAPDH was used as an internal control. (D). The protein expression of Wnt5a, FZD4, Active β-catenin, β-catenin, GSK3-β-p, Notch1, and GAPDH detected by Western blotting after 14 days of culture. (E). The statistical analysis of Western blotting. Data were given as the mean ± S.D. from three independent experiments. *P < 0.05, **P < 0.01. ***P < 0.001.

Abbreviations: CON, control; OM, osteoblast inducing conditional media; DEX, dexamethasone; DEX+ED-71, dexamethasone+eldecalcitol.