Figure 2.

Effects of WEPK on osteoclastogenesis in vitro: (A) BMMs were treated with WEPK (0–200 µg/mL) for 1 day, followed by assessment of cell viability. (B) MLO-Y4 cells were treated with or without VitD₃ (10 nM) for 1 day, following pre-treatment with WEPK (200 µg/mL) for 3 h. The mRNA expression of the target genes was assessed by real-time PCR. (C,D) BMMs and MLO-Y4 cells were cocultured for 5 days with VitD₃, either without RANKL (C) or with RANKL (50 ng/mL, (D)), following WEPK pre-treatment. TRAP staining, TRAP activity, and quantification of TRAP-positive osteoclasts (TRAP(+) MNCs) were performed. (E) BMMs were cultured for 3 days with M-CSF (60 ng/mL) and RANKL (50 ng/mL) after pre-treatment with WEPK. TRAP staining, TRAP activity, and quantification of TRAP(+) MNCs were performed. Scale bar, 200 μm, ** p < 0.01, *** p < 0.001 vs. control.