Fig. 5

C3 and C4 inhibit osteoclast differentiation and formation stimulated by IL-1β. Pre-osteoclasts were cultured in the presence of C3 (5 mmol/L) or C4 (1 mmol/L) with IL-1β (40 ng/ml) for 5 d. Mature osteoclasts were harvested for TRAP staining (a) and quantification of TRAP-positive multi-nucleated (≥ 5 nuclei) osteoclasts (b). Relative expression of mRNA (c) and proteins in osteoclast differentiation (d) were analysed. e Representative images of osteoclast immunofluorescence co-immunostaining with TRAF2 (green), NFATc-1 (red), and DAPI (blue). Results are mean ± SEM, ^**P < 0.01, ^***P < 0.001. C3 propionate, C4 butyrate, IL-1β interleukin 1 beta, RANKL receptor activator of NF-κB ligand, TRAF2 TNF receptor-associated factor 2, NFATc-1 nuclear factor of activated T-cells