Fig. 3.

Depletion of ROS with NAC or ebselen does not inhibit anti-IgM- or SDF-1-induced ERK 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 30 μg/ml anti-IgM or b 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 forms of ERK1 and ERK2 (upper panels). The blots were then stripped and reprobed with anti-ERK1/2 antibodies to show that equivalent amounts of ERK1/2 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