Transcriptomic profile of WT and Mthfd2 KO alloreactive CD4+ T cells [RNA-seq]

To investigate the effect of Mthfd2 KO in alloreactive CD4+ T cells. We performed in vitro mixed lymphocyte to induce alloreactive T cells. CD4+ T cells were respectively purified from the spleens of 8-week-old TEaWT and TEaMthfd2 KO mice by magnetic-activated cell sorting (MACS) positive selection. APCs were sorted from the spleens of 8-week-old F1 (BALB/c x C57BL/6) mice. CD4+ T cells were cocultured with APCs three days and then performed RNA-sequencing. CD4+ T cells were respectively purified from the spleens of 8-week-old TEaWT or TEaMthfd2 KO mice by magnetic-activated cell sorting (MACS) positive selection. APCs were sorted from the spleens of 8-week-old F1 (BALB/c x C57BL/6) mice and stained with mitomycin for 30 min at 37 °C with 5% CO2. Then, CD4+ T cells were then cocultured with APCs for 3 days at 37 °C and 5% CO2 in RPMI 1640 medium supplemented with 10 mM HEPES, 50 mM 2-mercaptoethanol, 100 U/mL penicillin/streptomycin, and 2 mM glutamine.