Figure 2.

Sclerostin inhibits Wnt3a-induced osteoblast differentiation without affecting canonical Wnt signaling in C3H10T1/2 cells. (a) ALP activity of C3H10T1/2 cells pre-incubated with 10 μg ml⁻¹ sclerostin, 1 μg ml⁻¹ dikkopf 1 (DKK1) or vehicle and stimulated with 20% control-conditioned medium or Wnt3a-conditioned medium for 3 days. (b) Evaluation of β-catenin transcriptional activity in C3H10T1/2 cells pre-incubated with 10 μg ml⁻¹ sclerostin, 1 μg ml⁻¹ dikkopf 1 or vehicle and stimulated with 20% control-conditioned medium or Wnt3a-conditioned medium for 24 h. (c) Western blot analyses of LRP5/6 phosphorylation in C3H10T1/2 cells stimulated with Wnt3a-conditioned medium for different incubation times; and analysis of LRP5/6 phosphorylation in C3H10T1/2 cells pre-incubated with 10 μg ml⁻¹ sclerostin, 1 μg ml⁻¹ dikkopf 1 or vehicle and stimulated with 200 ng ml⁻¹ recombinant Wnt3a or its vehicle for 3 h. *P<0.01 for Wnt3a versus control; +P<0.01 for SOST or DKK1 versus vehicle treatment.