Figure 4

CO inhibited FADD phosphorylation via blockage of JNK signaling. 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 Western blot analysis to detect phosphorylated and total FADD (A), or to immunoprecipitation (IP) with anti-Fas followed by immunoblotting to detect phospho-FADD (B). Lysates were subjected to immunoblotting to detect phospho- and total JNK. (C). MLEC cultures were treated with antibody Jo2 (200 ng/ml) for the indicated times, in the absence or presence of JNK inhibitor (20 μM). The total lysates were subjected to immunoprecipitation with anti-Fas followed by immunoblotting to detect phospho-FADD (D). Total FADD (A, B, D) or total JNK (C) served as the standards. Westerns are representative of three independent experiments.