Reduction of MCA by RANKL treatment is caused by decreased expression of ezrin. (A) Western blot analysis of cell lysates from RAW 264.7 cells without or with RANKL treatments for the indicated times, using anti-ezrin, anti–p-ERM, and anti–β-actin antibodies. (B and C) Quantification of (A). Mean ± SEM, n = 12 experiments. P value obtained from one-way ANOVA with Tukey’s test. **P < 0.01; ****P < 0.0001. (D) Confocal images of RAW 264.7 cells without or with RANKL for 60 or 84 h, stained with anti–p-ERM antibody (green) and DRAQ5 (gray). Scale bar: 20 μm. (E) Quantification of fluorescence intensity of p-ERM beneath the PM determined by wheat germ agglutinin (WGA) signal. Mean ± SEM, n = 40 cells for each condition. P value obtained from one-way ANOVA with Tukey’s test. ****P < 0.0001. (F) Confocal images of siRNA knockdown cells treated with RANKL for 70 h, stained with phalloidin (magenta) and DAPI (gray). Multinucleated cells are surrounded by dotted lines. Scale bar: 50 μm. (G) Quantification of fusion index in F. Dotted lines in violin plots show median and quantiles. n = 30 fields of view for each condition. P value obtained from one-way ANOVA with Tukey’s test. *P < 0.05; ****P < 0.0001. Source data are available for this figure: SourceData F3.