FIGURE 5.

Zbed3 could inhibit GSK3β phosphorylating β-catenin through binding Axin. Axin binding with β-catenin (A) or GSK3β (B) was not affected by Zbed3. Co-immunoprecipitation and Western blotting were done with transfected 293T cells as described above. C, Zbed3 inhibited GSK3 phosphorylating β-catenin. 293T cells were transfected with plasmids as indicated. 24 h later, the cells were lysed, and FLAG-tagged β-catenin was immunoprecipitated and checked with FLAG or phosphor-β-catenin antibody, respectively. D, Zbed3-SA or -TA could not affect the phosphorylation of β-catenin by GSK3β. Experiments were manipulated as in C with 293T cells. E, Wnt-3a CM-stimulated dephosphorylation of β-catenin was blocked by Zbed3 siRNA in NIH3T3 cells. 48 h after transfection of the indicated siRNA, Wnt-3a CM was added for additional 3 h with MG132 (25 μm), and total cell lysates were used for Western blotting assay with the indicated antibodies.