FIGURE 5.

Activation of Runx2 expression requires calycosin in DBT.(A) In pRunx2-Luc transfected osteoblasts, the promoter activity was revealed by luciferase assay in cultured osteoblasts by treating a series of DBT decoctions at different concentrations for 7 days. Dexamethasone plus vitamin C (Dex; 50 nM, Vit C; 250 μM) was used as positive control, which induced ∼4-fold activation. (B) Cultured osteoblasts were treated with a series of DBT (1.0 mg/mL) for 7 days. Cell lysates were collected to determine the protein expression of Runx2 (∼57 kDa) using specific antibody. Histone-1 (∼30 kDa) served as a loading control. Quantification of protein amount from the blot was calculated by a densitometer. Values were expressed as the ratio to the basal reading, where the control (untreated culture) equaled to 1 and in Mean ± SEM, where n = 3. ^∗∗∗p < 0.001, ^∗∗p < 0.01 as compared to the control.