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The brain tissue samples were homogenized in RIPA buffer containing protease inhibitor cocktail (Kangchen Bio-tech, Shanghai, China) and 1 mM Phenylmethylsulfonyl fluoride (PMSF), followed by 1 min of sonication. Samples were then centrifuged and the supernatant containing the whole tissue extract was retained. Protein concentration was determined using a BCA assay. Sample preparation was based on previous studies [57], and all samples were processed simultaneously. Samples were analyzed using LC-MS/MS.
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