Extended Data Fig. 7: pEtn-treated T cells have changes in metabolic and transcriptional pathways, and functional changes driven by pEtn can be rescued with PMA/Ionomycin.

A) Quantification of metabolites in TCA cycle, oxidative phosphorylation, glycolysis, glutaminolysis and pentose phosphate pathway of SIINFEKL-activated OT-I T cells in RPMI+pEtn versus RPMI culture B) Quantification of lipid metabolites of SIINFEKL-activated OT-I T cells in RPMI+pEtn versus RPMI culture as shown in Fig. 3a. C) Volcano plot of RNA-Sequencing of activated OT-I T cells cultured in RPMI or RPMI +pEtn. Highlighted genes represent exhaustion associated genes. D) Gene set enrichment assay of RNA-Seq data shown in C) of activated OT-I cells cultured in either RPMI or RPMI+pEtn. (E-G) OT-I T cells activated by SIINFEKL O/N were cultured in RPMI for 2 days and then in either RPMI or RPMI+pEtn for 5 days as described in Fig. 3a). Then the cells were restimulated by either α-CD3 and α-CD28 or PMA/Ionomycin for 6 hours in RPMI. After restimulation, cells were analyzed for E) IFNγ and TNF co-expression F) Granzyme B expression, and G) IL-2 expression by flow cytometry. Normalized MFI ±SEM is quantified for Granzyme B and IL-2, while %IFNγ⁺TNF⁺ cells ±SEM are quantified on the left of each plot from three independent experiments (n = 6 mice/group, p-values left to right E) p = 0.0002, p = 0.6170, F) p = 0.0046, p = 0.0267, G) p = 0.0007, p = 0.2566). two-tailed paired t-test. ns, non-significant. Source numerical data are available in source data.

Source data