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RESEARCH PRODUCT

Additional file 2: of Fibroblasts from bank voles inhabiting Chernobyl have increased resistance against oxidative and DNA stresses

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The repair efficiency of nicked, oxidized, or linear plasmids is similar in control and Chernobyl bank vole fibroblasts. For host-cell reactivation assay, 5000 cells were plated on 96 well plate, treated next day with 20 μM etoposide for 8 hours, and then transfected with pGL3 (Promega) plasmid treated either with Nb.BsmI that nicked the plasmid coding sequence three times, with HindIII that linearized the plasmid after promoter sequence, or with 50 μM FeSO4 and 1 mM H2O2, which created oxidative damage on the plasmid. To control transfection efficiency cells were transfected also with pNL1.1 nano-luc vector. Luciferase expression was analysed 24 h after transfection with Nano-Glo Dual-Luciferase reporter assay system as suggested by the manufacturer (Promega). The bar charts show the relation of standardized treated to standardized untreated plasmid expression. The results are from four separate experiments using the eight Chernobyl (N = 32) and eight control cell lines (N = 32). Variation is shown by standard deviation. (PDF 27 kb)