Repressed Blautia-acetate immunological axis underlies chronic stress promoted breast cancer progression

To explore acetate wether exerted its effects on T cell recall function and metabolism via transcriptional regulation. We used the Easy sepTM Mouse CD8+ T Cell Isolation Kit for negative selection of CD8+ T cells. Then CD8+ T cells were cultured in complete RPMI-1640 culture medium with recombinant mouse IL-2 (15 ng/ml) and anti-CD28 (1 µg/ml) in plate pre-coated with anti-CD3 (2 mg/ml). CD8+ T cells (2×106 cells/ml) were treated with or without sodium acetate (5 mM) for 24 hour, and co-cultured with/without 4T1 cells (1×105 cells/ml) for 48 hours. Subsequently, we removed the dead cells with EasySepTM Dead cell Removal (Annexin V) Kit and collected CD8+ T cells for RNA sequencing. Overall design: Comparative gene expression profiling analysis of RNA-seq data for CD8+ T cells after different treatments.