Extended Data Fig. 5: OTI CD8+ T cell proliferation induced by IL-7 and IL-12 requires recognition of self pMHC.
From: T cell receptor and cytokine signal integration in CD8+ T cells is mediated by the protein Themis
(A) WT OTI lymphocytes were incubated with 50ng/ml each IL-7 and IL-12, with or without 50ng/ml E1 peptide in the presence or absence of 10μg/ml CD8β blocking antibody for 7 days. The numbers shown denote mean +/- SEM from 3 mice from 1 experiment, representative of 2 independent experiments. (B) Acute Themis deletion reduces responses to IL-7 and IL-12. FACS-sorted naïve CD44low CD8+ T cells from tamoxifen-treated ERT2-Cre+ (Themis deletion) or ERT2-Cre- mice were Cell Trace Violet labelled and incubated in 50ng/ml IL-7 and IL-12 with or without 100 ng/ml E1 for 7 days. Cell proliferation was assessed by flow cytometry. Representative flow cytometry plots are shown, with numbers indicating mean +/- SEM of triplicates, obtained from 3-4 pooled lymphocytes/genotype. Data representative of two independent experiments. (C) WT or cKO OTI lymphocytes were incubated with 10ng/ml IL-7 and the indicated concentrations of IL-7 for 3 days. The percentage of divided cells were determined based on CTV dilution. Mean +/- SEM from 9 mice/genotype, pooled from 3 independent experiments.
