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Cultured cells were stimulated for 6 h with PMA (50 ng ml 1 ) (BioGems) and ionomycin (1 g ml 1 ) (BioGems) in the presence of brefeldin A (1 g ml 1 ) (GolgiPlug, BD Biosciences), stained for viability (Zombie UV Fixable Viability Dye (BioLegend)) and surface markers, fixed and permeabilized with 3.7% formaldehyde (Sigma) and 0.1% NP-40, respectively, and stained for intracellular cytokines

FDX1-mediated toxicity in cuproptosis occurs through the specific interaction of Cu+ with the mitochondrial lipoylase system, which triggers deleterious protein aggregation (173)

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Consequently, tumor cells exhibit a heightened affinity for Gln absorption within the TME, depleting the limited Gln supply and exerting an immunosuppressive effect (76)