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After 30 min at 4 C, cells were washed in perm/wash buffer and resuspended with secondary Ab anti-Rabbit IgG FITC (Fisher Scientific 15303926) or PE (CST 79408 S)
The brain was removed, then post-fixed with the fixative (4, overnight), and embedded in Tissue-Tek OCT-compound (Sakura Finetek, Tokyo, Japan)
The generation of ROS as by-products of the mitochondrial electron transport chain (ETC) involves superoxide radicals and hydrogen peroxide (Jomova et al
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