FIG. 7.

DISC analysis in wild-type (wt) and mutant Jurkat cells. Jurkat cells (wild type, FADD deficient [def], and caspase 8 deficient) were treated for 15 min with FasL, ACRP:FasL, or PBS alone, in the presence (+) or absence (−) of cross-linking anti-Flag M2 antibody, and were then washed in PBS. After lysis, samples treated without cross-linker were supplemented with anti-Flag antibody and ligands were immunoprecipitated and analyzed by Western blotting for Fas, FADD, and caspase 8. For each condition (except PBS alone), 1% of the postnuclear extract was loaded for comparison. The migration positions of procaspase 8 (Casp-8), FADD, and Fas and the high-molecular-mass modification of Fas (Fas hmw) are indicated by black arrowheads. The open arrowhead points to the heavy chain of the immunoprecipitating antibody (IgG).