Supplementary Figure 4: Ptpn2-deleted CD8⁺ T cells have increased proliferative capacity and in vivo persistence.

(a) Representative flow cytometry histogram of CTV dilution following restimulation of Slamf6⁺ control or Slamf6⁺ Ptpn2-deleted cells with αCD3/CD28 and IL-2. Vertical black lines separate peak numbers. Representative of two independent experiments, n = 3 mice. (b) Quantification of number of CD8⁺ T cells per division following in vitro restimulation (αCD3/CD28 with IL-2) of CTV-labeled Slamf6⁺ control or Slamf6⁺ Ptpn2-deleted CD8⁺ T cells isolated on day 8 post LCMV Clone 13 infection. Representative of two independent experiments, n = 3 mice. (c) Representative flow cytometry histogram of CTV dilution following restimulation of Tim-3⁺ control or Tim-3⁺ Ptpn2-deleted cells with αCD3/CD28 and IL-2. Vertical black lines separate peak numbers. Representative of two independent experiments, n = 5 mice. (d) Quantification of number of CD8⁺ T cells per division following in vitro restimulation (αCD3/CD28 with IL-2) of CTV-labeled Tim-3⁺ control or Tim-3⁺ Ptpn2-deleted CD8⁺ T cells isolated on day 8 post LCMV Clone 13 infection. Representative of two independent experiments, n = 5 mice. (e) Quantification of number of recovered Tim-3⁺ CD8⁺ T cells in the liver day 6 post LCMV Clone 13 infection following transfer of control or Ptpn2-deleted Tim-3⁺ CD8⁺ T cells isolated on day 8 post LCMV Clone 13 infection. Representative of two pooled experiments, n = 8 mice. (f) Frequency of dead cells of transferred Tim-3⁺ control or Tim-3⁺ Ptpn2-deleted cells day 6 post LCMV Clone 13 infection as in (e). Representative of two pooled experiments, n = 8 mice. Bar graphs represent mean and error bars represent standard deviation. Statistical significance was assessed by two-sided Student’s paired t-test (b, d), two-sided Student’s unpaired t-test (e), or two-way ANOVA (f) (ns p>.05, * p≤.05, ** p≤.01, *** p≤.001, **** p≤.0001).