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For metabolic flux assays, CD4 + T cells were washed with Seahorse XF RPMI 1640 media (Agilent Seahorse XF RPMI Medium with 1 mM HEPES (Agilent, Santa Clara, CA, cat#103,576100) with 10 mM Seahorse XF Glucose (Agilent, cat#103577-100), 1 mM Seahorse XF Pyruvate (Agilent, cat#103578-100), and 2 mM Seahorse XF L-Glutamine (Agilent, cat#103579-100) and plated at a density of 250,000 cells/well (50 L) in a Seahorse XFe96 plate (Agilent) pre-coated with Cell-Tak (Corning, Corning, NY)

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