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For short term treatments, seedlings at the age of 3 days post germination (DPG), were transferred, in darkness, to liquid MS supplemented with 20 nM, 40 nM methotrexate (MTX) or with 0.4% dimethylsulfoxide (DMSO) as a control

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This process is driven by disrupted GSH homeostasis, resulting from diminished GSH synthesis and impaired activity of two critical antioxidant systems: (1) the cystine/glutamate antiporter xCT, which imports cystine to fuel GSH production and (2) GPX4, which utilizes GSH to convert lipid peroxides into less toxic lipid alcohols (Stockwell et al., 2017