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 M [ 13 C 3 , 15 N]-cysteine-NEM, 36.4 M [ 13 C 2 , 15 N]-GSH-NEM, 10 M [D 4 ]-Cystine, 0.92 M [ 13 C 5 , 15 N 2 ]-GSSG, 20 M [D 4 ]-Hypotaurine, and 20 M of [ 13 C 2 ]-Taurine at a final concentration of 50 mg/mL followed by incubation on 4 C for 24 hr
It exists in both oxidized (NAD + ) and reduced (NADH) forms, the latter is generated by NAD + accepting high-energy electrons from glycolytic and TCA intermediates and acts as a primary electron donor in ATP synthesis to drive mitochondrial OXPHOS 518
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