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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Int J Oncol. 2011 Jan;38(1):249–256.

Figure 2.

Figure 2

Akt catalytic activity during TRAIL treatment. (A) DU-145 cells were treated with 200 ng/ml TRAIL for 4 h. Cells were lysed, and lysates were immunoprecipitated (IP) with mouse anti-Akt antibody. Catalytic activity of immunoprecipitated Akt after TRAIL treatment was estimated for GST-Bad or GST-GSK-3 fusion protein substrates. GST-Bad, phosphorylated GST-Bad, GST-GSK-3 and phosphorylated GST-GSK-3 were detected with anti-Bad, anti-phospho-Ser-136-Bad, anti-GSK-3α, or anti-phospho-GSK-3α/β antibodies, respectively. Immunoprecipitated Akt was detected with rabbit anti-Akt antibody. (B) DU-145 cells were lysed after exposure to TRAIL (200 ng/ml) for various times, and lysates were immunoprecipitated with anti-Akt antibody. Akt catalytic activity was examined in vitro using GST-Bad protein as a substrate. GST-Bad and phosphorylated GST-Bad were detected with anti-Bad and anti-phospho-Ser-136-Bad antibodies, respectively. Immunoprecipitated Akt was detected with rabbit anti-Akt antibody.