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. 1999 Aug 17;96(17):9683–9688. doi: 10.1073/pnas.96.17.9683

Figure 3.

Figure 3

Endogenous Apaf-1 does not interact with Bcl-xL or the other prosurvival homologs. (A) Tests with Bcl-xL and procaspase-9. Lysates from 293T cells transfected with wt procaspase-9 FLAG (lane 1), procaspase-9 C287A FLAG (lane 2), FLAG Bcl-xL (lane 3), or Bcl-xL FLAG (lane 4) were immunoprecipitated by using monoclonal anti-FLAG M2, fractionated by SDS/PAGE and then analyzed by Western blotting with monoclonal anti-Apaf-1 2E12. (B) Tests with other prosurvival Bcl-2 homologs. Lysates from 293T cells transfected with constructs expressing Flag-tagged mutant procaspase-9, Bcl-2, Bcl-w, Mcl-1, A1, Boo, E1B19K, or BHRF-1 were immunoprecipitated with anti-FLAG M2, and the blot was probed with anti-Apaf-1 2E12 (Upper). Probing of the same blot (Lower) with anti-Flag M2 confirmed expression and immunoprecipitation of each of the transfected Bcl-2 homologs (open arrows). HC, heavy chain; LC, light chain; PG, protein G. Data shown are representative of four experiments.