Comparison of IL10-eGFP+ cells with IL10-eGFP- CD4+ T cells induced by in vivo immunisation
Here we perform QuantSeq 3' mRNA sequencing of RNA extracted from flow sorted splenic CD4+ Il10-eGFP+ or CD4+Il10eGFP- T cells from Tg4 Nr4a3-Tocky Il10-eGFP mice 24 hours after immunisation with 4 mg/kg of [4Y]-MBP. Our aim was to confirm the Tr1 cell phenotype within the Il10-eGFP+ T cell subset Overall design: Tg4 Nr4a3-Tocky Il10-GFP mice were immunized through subcutaneous injection of 4 mg/kg [4Y] MBP peptide PBS into the flank. 24 hrs later mice were then euthanised and CD4+ Nr4a3-Timer+Il10-eGFP+ and CD4+Nr4a3-Timer+Il10-eGFP- T cells isolated and RNA extracted for 3' mRNA sequencing. 2 biological replicate mice were obtained for the 2 experimental groups. Each sample was run on 4 lanes, resulting in 48 FASTQ files which were then merged into single FASTQ files.