Figure 5. GSK3β associates with TRAF6, TAK1 and TRIF.

(a) BMDMs were stimulated with 10 μg ml⁻¹ poly I:C for 10 min and subjected to immunoprecipitation with an anti-GSK3β antibody. TRAF6 and TAK1 protein levels from whole-cell lysates (WCL) and TLR3 immunocomplexes (IP: α-GSK3β) were determined by western blotting. (b) GSK3β formed a ternary complex with TRAF6 and TAK1. HEK293T cells were transfected with the indicated combinations of expression plasmids. Co-imunoprecipitations were performed with anti-Flag antibody followed by western blotting with the indicated antibodies. The expression levels of the transfected plasmids were confirmed by western blot analysis of whole-cell lysates. (c) As in Fig. 4b, except that whole-cell lysates were immunoprecipitated with anti-TRAF6 antibody followed by western blotting as indicated. (d) GSK3β associated with TAB1 and TAB2 through TAK1. These experiments were performed as described in b. (e) TRAF6 associated with TAB1, TAB2, TAK1 and GSK3β. The experiments were performed as described in b. (f) GSK3β associates with TRIF, TRAF6 and TAK1. The experiments were performed as described in b. Data are representative of two or three independent experiments.