Figure 2.

Proteasome inhibition accumulates Bax to mitochondria/cytoplasm and increases interaction between Bax and Bcl-2 proteins. Jurkat cells (0 h; A and B) or Jurkat cells overexpressing Bcl-2 (0 h; C and D) were treated by either 10 μM lactacystin (A–C), 50 μM LLnV (D), or 50 μM LLM (A and B) for the indicated h. (A and B) Western blotting with antibodies to PARP and Bax were performed as described in Fig. 1. (C) Immunohistochemistry (see Materials and Methods). Localization of Bax protein (green) and nuclei (red) are shown. (D) Bax immunoprecipitates (IP), prepared by an agarose-conjugated N-20 Bax antibody, were immunoblotted (IB) first with the 6A7 Bax antibody (Upper) and then reblotted with a Bcl-2 antibody (Lower).