Research Paper Volume 6, Issue 3 pp 231—245

Oxidative stress and antioxidant response in fibroblasts from Werner and Atypical Werner Syndromes

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Figure 5. Analysis of the main antioxidant enzyme activities in atypical Werner syndrome and Werner syndrome fibroblasts. (A) CuZnSOD activity and (B) MnSOD activity evaluated using the Superoxide dismutase assay kit (Cayman, Ann Arbor, MI, USA) based on reduction of the tetrazolium salt by superoxide to give rise the formazan which is measured at 460 nm. (C) Catalase activity was analysed spectrophoto-metrically after the formation of the purpald-formaldehyde adduct which is measured at 540 nm. (D) Total glutathione peroxidase activity determined by the Glutathione peroxidase assay kit (Cayman, Ann Arbor, MI, USA) based on the decrease of NADPH after coupling the glutathione reductase reaction that uses the NADPH to reduce the GSSG produced by the Gpx enzymes after reducing the cumene-hydroperoxide reagents. Results are represented as mean (±SD) of three replicates of each cell line.