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Protein preparation and Western blot analysis Cultured cells or flashfrozen tissues were homogenized in icecold radioimmunoprecipitation assay (RIPA) buffer (50 mM TrisHCl pH 7.6, 150 mM NaCl, 0.5% sodium deoxycholate, 1% NP40, 0.1% SDS, 2 mM EDTA) containing COMPLETE protease inhibitor cocktail (Roche) and phosphatase inhibitors (Santa Cruz Biotechnology)

doi: 10.1056/NEJMoa2204705 177 MindellJ

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For a standard 150-pound adult, this equates to nearly 7 grams of each compound daily

This effect was counteracted by knockdown of Usp2 , while Usp2 OE further intensified the fluorescence, indicating a heightened level of lipid oxidation