Figure 2.

Inhibitory effects of phloretin on RANKL-induced actin ring formation (a), and induction and phosphorylation of gelsolin and vinculin (b, c, and d). Raw 264.7 macrophages were cultured in α-MEM for 5 days in the absence and presence of 1–20 μM phloretin. Ovariectomized (OVX) mice were orally treated with 10 mg/kg/day phloretin daily for 8 weeks. RANKL-differentiated cells were fixed in 4% paraformaldehyde for 10 min and rhodamine phalloidin was added to fixed cells (a). Fluorescent images were taken with a fluorescence microscope. Original magnification of microscopic images (n = 3), 400-fold. Cell lysates and bone tissue extracts were subject to SDS-PAGE and Western blot analysis with a primary antibody against gelsolin, vinculin, or phospho-vinculin (b, c, and d). Representative blot data were obtained from three independent experiments, and β-actin protein was used as an internal control. The bar graphs (mean ± SEM) in the bottom panels represent quantitative results of blots obtained from a densitometer. Respective values not sharing a common letter are different at P < 0.05.