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After 24 h, the medium was replaced with XF base medium (containing 1.25 mM sodium pyruvate, 25 mM glucose and 4 mM glutamine) 1 h before the assay

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Copper Drives Remodeling of Metabolic State and Progression of Clear Cell Renal Cell Carcinoma

L4524, Sigma-Aldrich, Taufkirchen, Germany), DMEM (Lonza), RNeasy kit (Qiagen), Dionex Ultimate 3000 UHPLC system (Thermo Scientific TM, Dionex, San Jose, CA, USA), penicillin/streptomycin (Dutscher), heat-inactivated FBS (Corning), PBS, RIPA, Griess reagent, NaNO 2 , loading buffer, 10% SDS-PAGE gel, PVDF membrane, anti-catalase, AF3398 from R&D Systems Noyal Chtillon sur Seiche, France, dilution 1/400

FASN uses acetyl-CoA as a substrate to synthesize saturated fatty acids (SFAs), whereas SCD1 can convert SFAs to MUFAs, thereby reducing the sensitivity of cells to ferroptosis [27,28,29]