TGF-β suppressed the differentiation, but not the effector functions of Tcf-1 ⁺ stem-like T cells. Similar experimental setup to Fig. 1. (A–C) Day 22 after infection, the percentage of IFN-γ⁺ (A), IL-2⁺ (B), and granzyme A⁺ (C) cells in Tcf-1⁺ and Tcf-1⁻ subsets of donor P14 T cells are shown. (D) Representative FACS profiles to show the expression of Tox. (E) MFI of Tox in Tcf-1⁺ and Tcf-1⁻ subsets are shown. (F) MFI of Tim-3 is shown. (G) Experimental setup for H–J. (H) The percentage of donor CD8⁺ T cells in the total CD8⁺ population is shown. (I) Representative FACS profiles of pre-gated donor CD8⁺ T cells are shown. (J) The percentage of Tcf-1⁺ (left) and CX3CR1⁺ (right) cells in donor CD8⁺ T cells are shown. Each pair of symbols in A–D, and each symbol in E, H, and J represents the results from an individual recipient mouse. Pooled results from two (A–C, n = 8–12; D and E, n = 5; H–J, n = 4/each) and five (F, n = 19) independent experiments are shown. **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 by ordinary one-way ANOVA (A–F) or Student’s t test (H–J).