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Mechanistically, glutamine is a major precursor for glutathione synthesis, therefore, limiting the utilization of glutamine by tumor cells leads to the reduction in GSH, inhibits the activity of sarcoplasmic reticulum Ca 2+ -ATPase (SERCA), and activates the NF-B signaling pathway, thereby promoting the expression of tumor PD-L1 and inactivating the co-cultured T cells

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After 96 hours, wells were aspirated, washed with PBS, and a fixative/staining solution containing 3.7% formaldehyde (Fisher Scientific F75P-1GAL) and 5 g/mL Hoechst 33342, Trihydrochloride, Trihydrate (Invitrogen H1399) was added