Splenocytes from chronically infected mice (>45 days p.i.) were stimulated with GP33-41 peptide for 5 h followed by phenotypic marker staining and intracellular staining. a, Gating strategy for IFN-γ+PD-1+CXCR5+Tim-3− and IFNγ+PD-1+CXCR5−Tim-3+ CD8+ T cells. b, Representative FACS plots and graph showing the frequency of TNFα- and IL-2-producing cells among IFN-γ+CD8+ T cells. Data are representative of 2 experiments (n = 4 per experiment). Student’s paired t-test, where **P < 0.01.