Figure 6. Wnt inhibition rescues kyphosis of Eed cKO mice.

(a) The effect of Wnt inhibition on kyphosis was assessed by the spinal cervical angle in P7.5-old mice. C59 Wnt inhibitor (Wnt-i) treatment from E14.5 through P7.5 significantly ameliorated kyphosis in Eed cKO mice. (b) Cervical angle measurements. *P<0.05 versus vehicle-treated Ctrl. **P<0.05 versus vehicle-treated cKO. Error bars show the s.e.m., unpaired Student's t-test was used. (c) Expression of active (non-phosphorylated) β-catenin of growth plate sections of Ctrl and cKO mice was assessed 6 h after C59 (Wnt-i) or vehicle injection. A significant reduction of active β-catenin protein was observed in C59-treated animals. Scale bar, 20 μm; red bar indicates the proliferating chondrocyte zone and blue bar indicates the hypertrophic zone. (d) In vitro treatment with C59 (Wnt-i) decreased the active (non-phosphorylated) β-catenin level in Eed cKO primary rib chondrocytes. Chondrocytes were cultured for 24 h in the presence of C59 (Wnt-i). (e) Col10a1 in situ hybridization on sagittal sections of the cervical spine of E17.5 embryos shows suppression of accelerated hypertrophic differentiation. C59 was injected into pregnant mothers from E 14.5 through E 17.5. C2, second cervical vertebra. Scale bar, 200 μm.