Extended Data Fig. 1: Characterization of chronically stimulated T cell exhaustion phenotype.
From: LARP4-mediated hypertranslation drives T cell dysfunction in tumors
(a) Bar plots showing the percentage of cells expressing IFN-γ, TNF, or IL-2 in acutely stimulated d8 TEFF cells, and chronically stimulated d4 and d8 TEX cells (n = 4). (b) Bar plots showing the levels of mitochondrial reactive oxygen species (mtROS, measured using MitoSox) in d8 TEFF, d4 TEX, and d8 TEX cells (n = 3). (c) Oxygen consumption rate (OCR) using the Seahorse XF bioanalyzer measuring the respiratory capacity of d8 TEFF cells, d4 TEX cells, and d8 TEX cells. Cells were seeded at 5 ×105/well in the XF96 plate (n = 5). (d) Representative flow cytometry analysis showing the expression of PD-1 and TIM-3 in d8 TEFF, d4 TEX, and d8 TEX cells. (e) Bar plots showing the MFI of PD-1 and TIM-3 in d0 TN, d8 TEFF, d4 TEX, and d8 TEX cells (n = 3). (f) Bar plots showing the percentage of PD-1hi TIM-3+ CD8+ T cells in d8 TEFF, d4 TEX, and d8 TEX cells (n = 4). (g) Flow cytometry gating strategy to identify PD-1hi TIM-3− early TEX and PD-1hi TIM-3+ terminal TEX in tumor-infiltrating OT-I T cells. n, number of technical replicates. For statistical analysis, figures (a-c and e-f) are presented as mean ± SEM and figures (a-b and e-f) were analyzed using unpaired two-sided Student’s t-tests. The data represent three independent experiments (a-c and e-f).
