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. 2002 May;4(3):218–227. doi: 10.1038/sj.neo.7900233

Figure 3.

Figure 3

Phospho Bcl2 has shorter half-life (t{1/2}) than that of native Bcl2 and proteasome inhibitors can stabilize phospho Bcl2 even in the presence of cycloheximide (CHX). Panels A and B: 16-hour 1 µM Taxol-treated W-34 cells were incubated with or without proteasome inhibitors (20 µM MG132/10 µM lactacystin) in the presence or absence of CHX (20 µg/ml) for the indicated periods. Panel A: 16-hour Taxol-treated cells were further incubated. Lane1: no CHX (0 hours); lanes 2 and 7: 2 hours without or with CHX treatment, respectively; lanes 3 and 8: 4 hours without or with CHX, respectively; lanes 4 and 9: coincubation for 4 hours with either MG132 (lane 4) or CHX and MG132 (lane 9); lanes 5 and 10: 6 hours without or with CHX, respectively; lanes 6 and 11: coincubation for 6 hours with either MG132 (lane 6) or CHX and MG132 (lane 11). Panel B represents identical experiments except another proteasome inhibitor lactacystin was used instead of MG132. Panel C: W-34 cells were incubated with or without 20 µg/ml CHX for indicated time periods. Equal amounts of proteins were subjected to immunoblotting using Bcl2 monoclonal antibody.