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Co-immunoprecipitation (CoIP) For immunoprecipitation assays, cells were resuspended in lysis buffer (50 mM Tris, pH 8.0, 150 mM NaCl, 50 mM EDTA, 1% NP-40, 10% glycerol) supplemented with 50 g/mL PMSF and 1 protease inhibitor cocktail (Cat No.20109ES05, Yeasen)

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Importantly, the recent 3D structure of human ASCT2 confirmed the results reported above (Garaeva et al., 4) representing a fundamental pre-requisite to inform important emerging studies in ASCT2 biology, i.e., interactions with drugs and xenobiotics (see section Involvement in Human Pathology and Interaction With Drugs)