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Stained cells were then analyzed with a Verse flow cytometer and BD FACS Diva software (BD Biosciences, San Jose, CA)

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In general, the rate of synthesis, regeneration, and recycling of glutathione is determined by three key factors 2 : Cellular levels of cysteineCysteine availability is the rate-limiting step in glutathione production, so not enough cysteine and de novo synthesis of glutathione will slow Enzyme glutamate cysteine ligase (GCL) activity, in part regulated by glutathione feedback inhibition Glutathione depletion due to high levels of oxidative stress, inflammation, or exposure to xenobioticsin these situations, de novo synthesis is upregulated Of course, as an antioxidant, one of the most important roles of glutathione is shielding cells from reactive oxygen species and reactive nitrogen species, but thats not its only job