Figure 6.

Wnt1 suppresses osteoclastogenesis through canonical Wnt signaling pathway. (A, B) C3H10T1/2 control or Wnt1‐overexpressing cells (A) were plated on the bottom of cell culture wells and growth‐arrested by 2.5 μg/mL mitomycin C treatment for 2 hours. Raw264.7 cells were plated on top of them. Osteoclastogenesis was induced by 50 ng/mL RANKL. Cells were treated with DMSO, 10μM XAV939, 10 μM KN93, or 5μM SP600125. (B) At D5 the cultures were stained for TRAP to demonstrate mature multinucleated osteoclasts. (C–H) mRNA expression of osteoclast‐specific and Wnt target genes was measured by qRT‐PCR. All cell culture experiments were repeated three independent times with at least three replicate wells per condition. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001. Statistical analysis was performed by Student's t test.