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After 2 h incubation, cells were washed twice with cold PBS and were extracted with 80% ice-cold methanol at 2 × 10 6 cells/mL
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These alterations result from generation of highly reactive free radicals leading to peroxidation of membrane phospolypids rich in polyunsaturated fatty acids, particularly in conditions with increased intracellular iron and decreased antioxidant systems (namely glutathione) (Hirschhorn and Stockwell, 2019)
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