Fig. 1

The effect of betulinic acid (BetA) on cell viability and osteogenic differentiation in MC3T3-E1 preosteoblast cells. a Cell viability assay. The MC3T3-E1 cells were cultured with different concentrations of BetA (0 μM to 30 μM) for 24 h, and cell viability was determined by WST-1. b ALP staining. The MC3T3-E1 cells were cultured in osteogenic medium containing different concentrations of BetA (0 μM to 20 μM) for 72 h, and then ALP staining was performed as in Materials and Methods. c RT-PCR analysis. The MC3T3-E1 cells were cultured as in (b). Total RNA was isolated and RT-PCR was performed with specific primers for ALP, BMP2, BSP and OC. The level of GAPDH was examined as a loading control. Representative data are shown. ALP, alkaline phosphatase. BSP, bone sialoprotein. OC, osteocalcin. n = 3