Inhibition of osteoclastogenesis by canonical Wnt signaling. (A) Quantification of TRAP-positive multinucleated cells generated in the presence of different concentrations of LiCl, Wnt3a, or Wnt5a as indicated. (B) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture in the permanent presence of recombinant Wnt3a and Rankl as indicated. (C) Quantification of TRAP-positive multinucleated cells generated after 7 d of culture, when Wnt3a (or LiCl) and Rankl were added from day 4 of culture. (D) qRT-PCR expression analysis for the indicated genes in bone marrow cultures at day 3 of differentiation after treatment with 200 ng/ml Wnt3a for 6 h. All error bars represent mean ± SD (n = 6). Asterisks indicate statistically significant differences compared with controls. (E) Western blot analysis for phosphorylated Lrp6 and β-catenin after stimulation of bone marrow cultures at day 3 of differentiation by Wnt3a in the absence or presence of the Fzd8-CRD. The black arrows indicate the position and molecular weight of the nearest marker.