Fig. 5.

ACSC-derived TNFAIP3 suppresses osteoclastogenesis in vitro. a TRAP staining (top) and immunofluorescence staining of TRAF6 (middle) and CTSK (bottom) showed that the culture supernatant of ACSCs (10%) and soluble TNFAIP3 protein (20 ng/mL) inhibit OC formation and OC differentiation-related gene expression, while addition of the anti-TNFAIP3 neutralizing antibody (100 ng/mL) promotes OC formation. Scale bars, 50 μm. b Quantification of TRAP⁺ multinucleated osteoclasts (> 3 nuclei) (left) and relative fluorescence intensity of TRAF6 (middle) and CTSK (right) in (a). The statistical significance of differences was determined using one-way ANOVA with multiple comparison tests (LSD). Experiments were repeated independently for more than three times. All data are shown as the mean ± S.D. **P < 0.01, ***P < 0.001; ns, not significant