Next Generation Sequencing analysis of IL-2 induced OT-I wild-type and OT-I Themis cKO cells for three days.

Thymocyte-expressed molecule involved in selection (Themis) was identified as a T cell-specific protein for T cell development. Recently, Themis was found to be required for T cell homeostasis by orchestrating TCR signaling and cytokine, especially γc family cytokine IL-7 signaling in peraphrial CD8+ T cells. Here, our study identifies Themis as a regulator for another γc family cytokine IL-2-induced proliferation. Transcriptomics and metabolomics reveal that Themis regulates Jak-Stat, Akt-mTOR as well as the downstream metabolic pathways. Moreover, how Themis regulates Shp1 phosphatase activity is still controversial. We further demonstrate that Themis negatively regulates Shp1 phosphatase activity on Jak1 and Jak3 to facilitate downstream signaling and subsequently metabolic reprograming, highlighting the importance of Themis in metabolic responses of CD8+ T cells. Fresh enriched naïve OT-I wild-type and OT-I Themis cKO cells were cultured with IL-2 (1μg/ml) for three days. mRNA profiles of the culturing cells were generated by deep sequencing in triplicate.