Effect of GSTP ablation on protein S-glutathionylation responses to ROS or RNS.

<p>(A) Crude BMCs and (B) BMDDCs derived from WT or <i>Gstp1/p2</i>^−/− mice were exposed to various concentrations of H₂O₂ (1, 5 or 10 mM) (left panel) or PABA/NO (0.5, 1 or 2 µM) (right panel), at 37°C for 15 min. Proteins were separated by non-reducing SDS-PAGE and protein S-glutathionylation evaluated by immunoblot with mouse anti-GSH antibodies. Even loading of protein was confirmed by probing with rabbit anti-Cdk2 antibodies. Goat anti-mouse and goat anti-rabbit fluorescent secondary antibodies were used and immunoblots imaged with a two-channel IR fluorescent Odyssey CLx imaging system. S-glutathionylation levels were quantified and normalized relative to Cdk2 loading for each concentration point. The ratio from no drug treatment was assigned a value of one and all other ratios are shown relative to this value. Bars represent the means (<i>±SD</i>) from three independent experiments.