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7-Dichlorofluorescin Diacetate (DCFDA) Assay for Production of Reactive Oxygen Species Cells were incubated in 10 M DCFDA (Sigma) in PBS at 37C for 30 min as previously described (Chiu et al., Quantification of 8-Hydroxy-2-Deoxyguanosine (8-OH-dG) as a Marker for Oxidative DNA Damage We used 8-Hydroxy-2-Deoxyguanosine (8-OH-dG) as a biomarker for oxidative DNA on brain cells exposed to different treatments using the commercially available HT 8-OH-dG ELISA II kit (R&D Systems) as guided by the manufacturers instructions

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Timing: When is the intervention most effective