Figure 3

B355252 partially restores glutamate-induced depletion of GSH and attenuates the increases in AIF and Bax expression in HT-22 cells. HT-22 cells were pretreated with B355252 for 2Â h prior to glutamate addition for 10Â h as described in the Methods. The level of GSH was measured in the cellular lysates with monochlorobimane (MCB) and glutathione-S-transferase (GST). AIF and Bax protein levels were determined by immunoblot with anti AIF and anti Bax antibodies. GAPDH was used to normalize for variations in protein loading. (A), Expression of GHS (B), Immunoblots of AIF and Bax. The bars depict densitometric analyses of the relative values of AIF (top bar graph) and Bax (lower bar graph) present in glutamate treated, glutamate and B355252 treated, and control cells (***p<0.001 compared to glutamate-treated cells).