Fig. 5

Loss of GSH content in microglia influences oxidative stress but not cell viability. EOC13.31 microglia cells were divided into six groups: either untreated (control) or pretreated with 500 μM NAC or 200 μM BSO for 18 h, prior to 22 mM EtOH exposure for 6 h. Confocal images (20×) of cells labeled with CellROX Deep Red Reagent to detect ROS generation (a). Semi-quantification analysis of ROS fluorescence intensity (b). Intracellular GSH levels were determined fluorometrically by labeling cells with 50 μM Monochlorobimane (c). Western blot analysis for Nrf-2 expression and nuclear translocation, where Lamin B1 and β actin was used as a loading control for nuclear and cytosolic fraction, respectively (d). Effect of the above treatment regimen on cell viability (MTS assay) (e). Values are presented as mean average ± SEM three independent experiments. Student-Newman-Keuls comparison analysis was used (*p ≤ 0.05; **p ≤ 0.005)