Fig. 5.

Bergenin inhibits RANKL-induced osteoclast differentiation in vitro. (A) Cytotoxic effects of Ber in BMMs after treatment for 24 h, 48 h, 72 h and 96 h. (B) mRNA expression of osteoclast marker genes (NFATC1, C-FOS and CTSK) were detected by RT-qpcr. (C) Western blotting performed that Ber suppressed the activation of osteoclast related protein (NFATC1, C-FOS and CTSK). (D) The density of the western blot bands in (C) were quantified by ImageJ. (E) F-actin ring and TRAP staining results for BMMs cultured with M-CSF (30 ng/ml), RANKL (50 ng/ml), and different concentrations of Ber (0, 10 and 20 μM) for 5–7 days. (F) The TRAP-positive area of osteoclasts was quantified per well. (G) The area of osteoclasts F-actin ring was quantified. The values are mean ± SD for 3 separate experiments. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.