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TEV protease was added for overnight cleavage

A previous study has shown that perturbation of this pathway decreases sensitivity to the chemotherapeutic methotrexate and increasing flux through this pathway has been proposed to increase methotrexate efficacy in patients (Kanarek et al., 2018)

The most direct consequence is the depletion of antioxidants like glutathione (GSH), antioxidant enzymes including glutathione peroxidase, gamma-glutamylcysteine ligase (-GCL), and SOD1, or related proteins such as nuclear factor erythroid-2-related factor 2 (Nrf2) [26, 61, 68, 90, 91] (Fig