Figure 1

GR knockdown-induced alterations in GR expression, GR activity, protein carbonylation and intracellular GSH/GSSG ratio in CL1-0 and CL1-0ΔGR cells. (A) CL1-0 cells and CL1-0∆GR cells grown overnight and the expression of glutathione reductase in these 2 cell lines were monitored by immunoblotting. Beta-tubulin was used as loading control in this study. (B) GR activity assays of CL1-0 cells and CL1-0∆GR cells were performed. Activity is reported as units per milligram of protein in the total cell extract. One unit of enzyme activity is equal to 1 μmol DTNB reduced/min at 25°Cat pH 7.5. (C) Oxidative carbonylation of proteins in CL1-0 cells and CL1-0∆GR cells were measured by colorimetric-based ELISA analysis. (D) Reduced and oxidized glutathione ratio in CL1-0 cells and CL1-0∆GR cells were measured by luminescence-based ELISA analysis. Values are the average of 3 independent measurements +/- the standard deviation (p < 0.05*, p < 0.01 ** and p < 0.001 ***).