FIGURE 5.

MDP treatment induces rapid dissociation of the Nod2·Hsp90 complex. A, RAW264.7 cells were treated with MDP for the indicated times. B, cells were pretreated with IFN-γ for 6 h and then stimulated with MDP for the indicated times. Cell lysates were immunoprecipitated (IP) with anti-Hsp90 antibody, and associated Nod2 was analyzed by Western blotting. C, the expression vectors for Nod2 deletion mutants used in the study. D, HEK293T cells were transfected with the indicated plasmids. Forty-eight hours after transfection, total cellular proteins were extracted. Mutant Nod2 proteins in the lysates were immunoprecipitated, and associated endogenous Hsp90 protein was analyzed by Western blotting. Data are representative of three independent experiments with similar results. S.E., short exposure; L.E., long exposure; n.s., nonspecific; E.V., empty vector.