Fig. 6.

Overexpression of TNFAIP3 in ACSCs inhibits osteoclastogenesis in vitro. a, b TRAP staining (top) and immunofluorescence of TRAF6 (middle) and CTSK (bottom) in osteoclasts treated with 10% culture supernatant of transfected ACSCs. Knockdown of TNFAIP3 promotes OC formation, while overexpression of TNFAIP3 inhibits OC formation. Scale bars, 50 μm. c Quantification of TRAP⁺ multinucleated osteoclasts (> 3 nuclei) (left) and the relative fluorescence intensity of TRAF6 (middle) and CTSK (right) in (a–b). 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 < 0.05, **P < 0.01, ***P < 0.001