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Western immunoblot analyses For the preparation of total liver homogenates, frozen liver pieces were homogenized in RIPA buffer (150 mM NaCl, 1% TritonX-100, 0.25% sodium deoxycholate, 0.1% SDS, 50 mM Tris, 1 mM EDTA, 1 mM PMSF, protease inhibitor cocktail, pH 7.4) on ice with a tissue tearor (BioSpec Products, Bartlesville)

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By age 35-40, most people have lost a significant portion of their peak glutathione levels even without these additional stressors