A purine metabolic checkpoint that prevents autoimmunity and autoinflammation (T cell RNA Seq dataset)

In this study, we investigate how FAMIN activity in dendritic cells determines the release of mediators that affect priming, activation and gene transcription of CD8+ T cells. Overall design: Supernatant from Famin-/- and Famin+/+ splenic CD11c+ dendritic cells were harvested following overnight incubation. CD8+ T cells were isolated from spleens and lymph nodes of OT-I;Rag–/– mice, stimulated with anti-CD3 and CD28 in the presence of supernatant obtained from Famin-/- or Famin+/+ dendritic cells for 24 hours. Total RNA was extracted using the RNeasy Mini Kit (QIAGEN, 74104) in accordance with the manufacturer's instructions. Libraries were prepared using TruSeq stranded mRNA library prep kit (Illumina, 20020594). Sequencing of libraries was performed using an Illumina NextSeq 500 platform with NextSeq 500-Mid Output kit generating 2x150bp end reads. FastQ files were quality-checked and any residual adaptor sequences were removed using TrimGalore. Reads were then aligned to the reference genome Ensembl Mus_musculus.GRCm38 using STAR. Differential gene expression analysis was performed on read count files using edgeR.