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NIK phosphorylates IκBα in the presence but not in the absence of rabbit reticulocyte lysate. In vitro-translated 35S-labeled F-NIK and IκBα in reticulocyte lysate or, after immunoprecipitation from reticulocyte lysate (IPIκBα + IPF-NIK), were incubated in a standard kinase buffer containing 2.5 μM okadaic acid, and samples were then analyzed by SDS/PAGE and autoradiography.
