Fig. 3

TCG inhibited osteoclastogenesis through suppressing Rap1 signaling pathway. A, B KEGG and GO analysis of differentially expressed genes between replicative BMMS treated with vehicle or 1 µM TCG for 3 days under RANKL and M-CSF stimulation. C GSEA analysis of Rap1 signaling pathway-related genes. D BMMs were stimulated with RANKL and M-CSF and simultaneously treated with or without 1 µM TCG for 15 min. Active Rap1 was detected using an active Rap1 detection kit. E BMMs were stimulated with RANKL and M-CSF and indicated doses of TCG for 3 days. Western blot measured the protein expression of integrin αv, integrin β3, c-Src and Pyk2. F BMMs were stimulated with RANKL and M-CSF and simultaneously treated with or without 1 µM TCG for an indicated period of time. Western blot assessed phosphorylated c-Src and Pyk2 protein expression. The relative protein expression statistics were presented in Additional file 1: Fig. S4A–F. G BMMs were induced to osteoclasts while simultaneously treated with indicated doses of TCG. Phalloidin staining showed the effect of indicated doses of TCG on F-actin ring formation (Scale bar = 100 µm)