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. 1997 Nov 25;94(24):12829–12832. doi: 10.1073/pnas.94.24.12829

Figure 3.

Figure 3

IL-1RAcP interacts with IRAK. (A) IL-1-dependent coprecipitation of IRAK with IL-1RAcP in 293IL-1RI and HeLa cells. Protein extracts from 293IL-1RI or HeLa cells untreated or treated with IL-1 (100 ng/ml for 5 min) were immunoprecipitated with antiserum to the extracellular domain of IL-1RAcP or IL-1RI. The immunocomplexes were analyzed by immunoblotting to detect IRAK. (B) Interaction of IRAK with transiently expressed IL-1RAcP. IL-1RI or Flag epitope-tagged IL-1RAcP was expressed in 293 cells by transient transfection and immunoprecipitated with antiserum to IL-1RI or with anti-Flag antibody, respectively. Anti-Flag antibody was used for immunoprecipitation of extracts of cells transfected with the vector alone. After separation by SDS/PAGE, the immunocomplexes were blotted with antiserum to IRAK. Reactive proteins were detected with horseradish peroxide conjugated to protein A. (C) Interaction of IRAK with the cytoplasmic domain of IL-1RAcP. Flag-tagged IL-1RAcP or AcP(1–403) transiently expressed in 293 cells was immunoprecipitated with anti-Flag antibody and immunoblotted to detect coprecipitating IRAK.