CD44 and CD62L expression in Tim-3– and PD-1–expressing TILs. TILs were harvested from CT26 tumor-bearing mice and stained with 7AAD to exclude dead cells and antibodies against CD8, CD44, CD62L, Tim-3, and PD-1. (A) Representative staining on CD8⁺ Tim-3⁻PD-1⁻, Tim-3⁻PD-1⁺, and Tim-3⁺PD-1⁺ TILs is shown. FMO, controls for CD44 and CD62L staining are shown. Data are representative of three independent analyses. (B) Summary data showing the frequency of effector/memory (CD44^hiCD62L^low) and central memory (CD44^hiCD62L^hi) and CD44^int cells within the CD8⁺ Tim-3⁻PD-1⁻, Tim-3⁻PD-1⁺, and Tim-3⁺PD-1⁺ TILs. *, P < 0.05; **, P < 0.01; ***, P < 0.001, one-way ANOVA and Tukey’s multiple comparison test. n = 3. Error bars represent SEM.