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In terms of experimental design and in order to limit possible confounding effects due to variability between litters or genetic differences across strains, experiments were designed to include animals from multiple litters, typically three independent litters per group and littermate WT animals used as controls 63

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Treatment of B6 mice with the GLS1 inhibitor BPTES significantly reduced histological scores in the spinal cord of affected animals, lowered the number of CD4 + T cells, IL-17A and IFN-producing CD4 + T cells in the spinal cord, substantially decreased clinical scores and weight loss, and ameliorated experimental autoimmune encephalomyelitis ( + CD4 - CD8 - cells), the urinary albumin/creatinine ratio, glomerular nephropathy score, and T-cell infiltration with IL-17A in the kidneys, relieving the EAE disease activity ( + T cells by inhibiting GLS1, glycolysis, and CD4 + T-cell differentiation in vitro by reducing the level of HIF1 protein, a key metabolic sensor in Th17 cells ( Table 1 )

Frosty container, placed at 80 C overnight to complete cryopreservation, and submitted to Active Motif for ATAC-sequencing

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