FIGURE 4.

IVA inhibits RANKL‐induced osteoclastogenesis via G_i‐coupled GPCR and AMPK. A‐C, E, F, Mouse BMDMs were stimulated with IVA (200 μM) in the absence or presence of H89 (1 nM or 1 μM) (A), the G_i inhibitor PTX (10 ng/mL) (B, C), compound C (0.1 μM) (E) under OC differentiation conditions (30 ng/mL M‐CSF plus 100 ng/mL RANKL) for 5 days. With TRAP staining, TRAP⁺ MNCs (>3 nuclei or >5 nuclei) were counted and considered to be OCs (C, F). D, BMDMs stimulated with IVA (200 μM) during each time point in OC differentiation condition media. Extracted proteins were separated by 10% SDS‐PAGE, and levels of phosphorylated‐AMPK and AMPK were determined by Western blot. Data are representative of at least three independent experiments (A, B, D, E). Data are presented as the mean ± SEM (n = 3 for C, F). ^* P < .05, ^** P < .01, ^*** P < .001. NS: not significant. Scale bar, 2 μm (A, B, E)