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(Using WT's result as a benchmark value) Experimental Conditions Cell Line: HT1080 cells (wild-type or xCT-knockout or xCT- overexpressed) Cystine Uptake Assay Kit Pretreatment: PBS (+) (cystine-free, 0.1% glucose, 0.01% Mg 2+ , 0.01% Ca 2+ ), 37 , 5 min Uptake Conditions: Cystine Analog / PBS(+) (cystine-free, 0.1% glucose, 0.01% Mg 2+ , 0.01% Ca 2+ ), 37, 30 min Radio Isotope Method Pretreatment: None Uptake Conditions: 0.05 mmol/l cystine + [ 14 C]cystine (7.4 kBq/ml) /PBS(+) (0.1% glucose, 0.01% Mg 2+ , 0.01% Ca 2+ ), 37, 2 min Application Data 3: Evaluation of intracellular uptake and redox level after treating with Erastin Erastin is a known inducer of ferroptosis

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