14-3-3σ interacts with GSK-3β.

<p>(A) CCE, R1 and D3 cells transfected with control or 14-3-3σ vectors were lysed and 14-3-3σ complex was pulled down by immunoprecipitation (IP) with a Flag antibody. Proteins in the complex were analyzed by Western blotting (WB). (B) CCE cells were transfected with each isoform vector. 14-3-3 complex was pulled down and GSK-3β was determined by Western blotting. (C) Association of 14-3-3σ with WT or mutant GSK-3β was determined by immunoprecipitation (IP) with a Flag Antibody followed by Western blotting (WB) with GSK-3β and p-GSK-3β antibodies. (D) CCE cells transfected with Flag-tagged 14-3-3σ (σ) or control (CTR) vectors were treated with wortmannin. 14-3-3σ complex was immunoprecipitated with a Flag antibody and GSK-3β was analyzed by Western blotting. (E) 14-3-3σ overexpression reduced GSK-3β activity. Data represents mean ± s.d. (n = 3). *, p<0.05; **, p<0.01. (F) CCE cells were transfected with control or 14-3-3σ vectors for 42 h followed by treatment with wortmannin (Wort) for 6 h. Total and phosphorylated protein levels of GSK-3β and Akt were detected by Western blot analysis.