Fig. 1.

Depletion of ROS with NAC or ebselen inhibits anti-CD40-induced JNK activation but does not inhibit anti-IgM- or SDF-1-induced JNK activation. WEHI-231 cells were pretreated with 30 mM NAC for 2 h or with 20 μM ebselen for 1 h and then stimulated for the indicated times with a 10 μg/ml of the 1C10 anti-CD40 monoclonal antibody, b 30 μg/ml anti-IgM or c 100 ng/ml SDF-1. When cells were pretreated with ebselen, the control cells were incubated with an equivalent volume of DMSO. Cell extracts were separated by SDS-PAGE and analyzed by immunoblotting with a phospho-specific antibody that recognizes the activated form of JNK (upper panels). The blots were then stripped and reprobed with anti-JNK antibodies to show that equivalent amounts of JNK were present in each sample (lower panels). Molecular mass markers (in kDa) are shown to the left. For each panel, similar results were obtained in at least three independent experiments