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. 2002 Mar 26;99(7):4331–4336. doi: 10.1073/pnas.072088099

Figure 5.

Figure 5

Procaspase-8L selectively associates with the BAP31 complex. (A) Anti-Flag immunoprecipitates from H1299 cells cotransfected with the indicated expression constructs in the presence of 50 μM zVAD-fmk were analyzed by SDS/PAGE and Western blotting with anti-HA or anti-Nex domain antibodies. Note that only procaspase-8L-HA immunoprecipitated with BAP31-Flag. *, A nonspecific protein detected by anti-HA antibody in lysates. (B) Procaspase-8L induces loss of cell viability when ectopically expressed. H1299 cells were transfected with a luciferase expression construct and pcDNA3 vector expressing either procaspase-8/a-HA or -8L-HA, in the absence or presence of 50 μM zVAD-fmk, and luciferase activity was measured 24 h later. (C) Procaspase-8L is not detected in the Fas DISC. The DISC was immunoprecipitated following stimulation of H9 lymphocytes with anti-Apo-1/Fas (19) and the presence of procaspase-8L and procaspase-8 in the precipitates was assessed by immunoblotting as in Fig. 4C. IgG, Ig heavy chain. (D) Wt or Fadd-deficient mouse embryo fibroblasts (20) were infected with Ad5 dl520E1B− for 50 h and procaspase-8L cleavage was analyzed as in Fig. 4A.