Antigen calibration of T Cell Receptor signalling strength and T cell differentiation in vivo

Here we perform QuantSeq 3' mRNA sequencing of RNA extracted from flow sorted splenic T cell from Tg4 Nr4a3-Tocky Tiger mice immunised with a high or low dose of MBP 4Y peptide, enabling identification of time and dose dependent transcriptional signatures Overall design: There are 7 experimental groups in biological triplicate from Tg4 Nr4a3-Tocky Tiger mice: 1) Control group, unimmuinsed splenic T cells. 2) 0.8ug 4Y MBP peptide immunised T cells isolated at 4hrs. 3) 0.8ug 4Y MBP peptide immunised T cells isolated at 12hrs. 4) 0.8ug 4Y MBP peptide immunised T cells isolated at 24hrs. 5) 80ug 4Y MBP peptide immunised T cells isolated at 4hrs. 6) 80ug 4Y MBP peptide immunised T cells isolated at 12hrs. 7) 80ug 4Y MBP peptide immunised T cells isolated at 24hrs