Interaction and phosphorylation of FADD by PKCζ.
(A) KG1a cellular extract was immunoprecipitated by anti-FADD monoclonal antibody, and immunocomplexes were analyzed by Western blot using anti-PKCζ monoclonal antibody. In parallel, cellular extract was analyzed by Western blot using anti-PKCζ monoclonal antibody. (B) FADD phosphorylation by PKCζ in KG1a and KG1a/G8 cells. After cell lysis, cellular extracts were immunoprecipitated by anti-PKCζ monoclonal antibody. Immunocomplexes were resuspended in buffer reaction with γ32P-ATP and FADD-agarose as substrate. FADD phosphorylation was analyzed as described in “Materials and methods.” In parallel, an aliquot of each sample was analyzed by Western blot using anti-PKCζ monoclonal antibody to quantify immunoprecipitated proteins.
