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They were treated with sildenafil, received 20 x 106 C26GM vaccine-primed splenocytes from H2d pIL-2/GFP mice, or were given both treatments as indicated. (B) The tumors were surgically removed 9 d later, and the CD8+ T cell infiltrate was determined by flow cytometry. The percentage of CD8+ T cells was plotted against the tumor size at the time of tumor harvest. The software-provided tool (Sigma plot) was used to fit a three-parameter exponential decay curve (y = 36.13 + 92xe5.93x). Pearson bivariate correlation, P = 0.0000002. Data are derived from three independent experiments, with each containing five mice per group. (C) Intratumoral single cell suspensions were labeled with anti-CD25 or -CD69 antibodies. Data are expressed as the percentage of positive cells gated on the CD8+ T cell population. (D) IL-2 production in the ACT groups is reported as the percentage of GFP+ CD8+ T cells. Data are derived from two independent experiments. The paired t test p-value is reported. (E) BALB/c mice were challenged with 0.5 x 106 C26GM cells on day 0. They were treated with sildenafil starting on day 0, the anti-CD8+ depleting antibody, or both. The one-way ANOVA p-values (PA) are indicated. Data are reported from one of two similar experiments. Error bar values are shown.

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