Figure 2

CO inhibited Jo2-dependent caspase-8 and Bax activation. MLEC cultures were pretreated in the absence or presence of CO (250 ppm) for 2 h prior to the addition of antibody Jo2 (200 ng/ml) for the indicated times. The total lysates were subjected to immunopreciptiation (IP) with anti-Fas, followed by immunoblotting (IB) to detect caspase-8. Total Fas served as the standard. The control lane (c) represents cell lysate taken immediately after Jo2 addtion (A). Lysates were subjected to Western blot analysis to detect caspase-8 (B). MLEC cultures were pretreated in the absence or presence of CO (250 ppm) for 2 h prior to the addition of antibody Jo2 (200 ng/ml) for the indicated times. Lysates were subjected to immunoprecipitation with antibody 6A7 that specifically recognizes the activated form of Bax, followed by immunoblotting with anti-Bax. Total Bax served as the standard (C). Westerns are representative of three independent experiments.