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doi: 10.1038/sj.onc.1204101 21 FaraonioRVergaraPDi MarzoDPierantoniMGNapolitanoMRussoTet al

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For the quantification of sulfur metabolites in the tissue samples, the tissue metabolites were extracted and derivatized with NEM in ice-cold extraction solvent (80% MeOH: 20% H 2 O containing 25 mM NEM and 10 mM ammonium formate, pH=7.0) which includes stable isotope labeled internal standards (20 μM [ 13 C 3 , 15 N]-cysteine-NEM, 36.4 μM [ 13 C 2 , 15 N]-GSH-NEM, 10 μM [D 4 ]-Cystine, 0.92 μM [ 13 C 5 , 15 N 2 ]-GSSG, 20 μM [D 4 ]-Hypotaurine, and 20 μM of [ 13 C 2 ]-Taurine at a final concentration of 50 mg/mL followed by incubation on 4 °C for 24 hr