Extended Data Fig. 1: Ablating Tle3 shows modest impact on the functions of antigen-responding CD8⁺ T cells without affecting viral clearance.

a. Experimental design. Naive P14⁺CD8⁺ T cells were adoptive transferred into CD45-disparate recipients at 2 × 10⁴ cells/recipient, followed by i.p. infection with LCMV-Arm and characterization of CD8⁺ T cells at the effector and memory stages. dpi, day post-infection. b. Gating strategy to identify CD45.2⁺ donor-derived P14⁺CD8⁺ T cells in CD45.1⁺ wild-type recipients. c. Detection of cytokine production in effector CD8⁺ T cells on 8 dpi. CD45.2⁺ wild-type or Tle3^–/– naive CD8⁺ T cells were transferred into CD45.1⁺ wild-type recipients, which were infected with LCMV-Arm the next day, following experimental design in a. GP33-induced production of IFN-γ (top) and TNF-α (bottom) was detected in CD45.2⁺CD8⁺ T cells. Representative contour plots (left) are from 2 independent experiments, with values denoting percentages of the gated population. Cumulative data (right) of the percentage of IFN-γ⁺ population in P14⁺CD8⁺ T cells and the percentage of TNF-α⁺ populations in IFN-γ⁺ P14⁺CD8⁺ T cells are means ± s.d. d. Detection of granzyme B expression in effector CD8⁺ T cells by intracellular staining on 8 dpi. Representative histographs (left) are from 2 independent experiments, with the values denoting geometric mean fluorescence intensity (gMFI) of granzyme B. Cumulative data (left) are means ± s.d. e–g. Detection of LCMV in recipient sera. The serum samples were collected from recipients of WT or Tle3^–/– P14⁺CD8⁺ T cells at the indicated time points, and LCMV was detected with either quantitative RT-PCR (e) or plaque assays (f, g). Note that in the plaque assay, sera were serially diluted for the measurement as exemplified for the 4 dpi samples (f), and plaque forming units (PFUs) at 1:10 dilution were plotted for all time points (g). n = 5 each for mice receiving WT and Tle3^–/– cells in f; data in e and g are means ± s.d. from two independent experiments. h. Detection of T_CM frequency at ≥30 dpi in the spleen (SP) and lymph nodes (LN) of CD45.1⁺ wild-type recipients where CD45.2⁺ Tle3^–/– and CD45.1⁺ CD45.2⁺ WT P14 cells were mixed for co-transfer, followed by LCMV-Arm infection. Donor-derived T_M cells were identified firstly by gating on CD45.2⁺ cells, in which CD45.1⁺ cells were from WT (black) while CD45.1^–GFP⁺ cells were from Tle3^–/– (blue). Bar graphs are cumulative data of frequencies of T_CM cells as means ± s.d. from two independent experiments. Statistical significance in c, d, e and h was determined with two-sided Student’s t-test. ns, not statistically significant; *, p < 0.05; ***, p < 0.001.