Extended Data Fig. 6: Sildenafil specifically acts on DCs.

a, Quantifications of TCF1⁺TIM3⁻ CD8⁺ T cells in the TME in each group (n = 6 vs 8 mice). b, Quantifications of CD44⁺PD-1⁺TIM3⁻CD8⁺ T cells, IFNγ⁺CD44⁺PD-1⁺CD8⁺ T cells, and CD44⁺PD-1⁺IFNγ⁺CD8⁺ T cells in the TME in each group (n = 6 vs 8 mice). c, Tumour growth and survival curves of B16F10^Pde5−/− bearing mice treated with 7 doses of vehicle or sildenafil every other day via subcutaneous (s.c.) injection (n = 10 vs 12 mice). d, Kaplan–Meier survival curve of previously cured mice rechallenged with a double dose of MC38^Pde5−/− cells (WT, n = 4; cured, n = 5). Age- and sex-matched mice served as controls. e, Quantifications of CD44⁺CD8⁺ and Ki67⁺CD8⁺ T cells in tdLNs from each group (n = 6). f, Representative flow cytometric histograms of IFNγ or GZMB in CD44⁺PD-1⁺CD8⁺ and quantifications of CD44⁺PD-1⁺CD8⁺ T cells and GZMB⁺CD44⁺PD-1⁺CD8⁺ T cells in the TME from each treatment group (n = 6 per group). GZMB, Granzyme B. g,h, TCGA pan-cancer analysis using DC gene signature combined with PDE5 expression (n = 1066 patients). Kaplan–Meier survival curve of patients stratified by median PDE5 expression into high or low (n = 537 vs 529 patients) DC gene signature groups (g). Correlation between PDE5 and CD8 T cell infiltration levels (h). i, Quantifications of mig-CD103⁺ and CD11b⁺ cDCs in Zbtb46^crePde5^fl/fl mice bearing MC38^Pde5−/− tumour and treated with vehicle or sildenafil (vehicle, n = 5; sildenafil, n = 4). j, Quantifications of tdLNs CD44⁺CD8⁺ or TME IFNγ⁺CD44⁺PD-1⁺CD8⁺ T cells in Zbtb46^crePde5^fl/fl mice bearing tumour and treated with vehicle or sildenafil (n = 5 vs 4 mice). k, Schematic of the design strategy for LSL-Pde5 mice. l, Survival curves corresponding to results in Fig. 5o. m, Quantifications of tdLN mig-CD103⁺ and CD11b⁺ cDCs in Zbtb46^crePde5^LSL mice bearing MC38^Pde5−/− tumour and treated with vehicle or sildenafil (n = 6 per group). n, Quantifications of tdLN CD44⁺CD8⁺ or IFNγ⁺CD44⁺PD-1⁺CD8⁺ T cells in Zbtb46^crePde5^LSL mice bearing MC38^Pde5−/− tumour and treated with vehicle or sildenafil (n = 6 per group). o, Tumour growth curves of Zbtb46^cre or Zbtb46^crePde5^LSL mice inoculated with MC38^Pde5−/− tumour. Data were re-analysed from Fig. 5o (Zbtb46^cre, n = 8; Zbtb46^crePde5^LSL, n = 7). p,q, Quantifications of tdLN migratory cDCs (p), or migratory CD103⁺ and CD11b⁺ cDCs (q) in Zbtb46^cre and Zbtb46^crePde5^LSL mice (n = 4 per group). r,s, Quantifications of CD44⁺CD8⁺ T cells in tdLNs (r) or IFNγ⁺CD44⁺PD-1⁺CD8⁺ T cells in the TME (s) in Zbtb46^cre and Zbtb46^crePde5^LSL mice (n = 4 per group). Data are representative (a, b, d, e-j, m, n, p-s) or merged (c, l, o) of at least two to three independent experiments and present as mean ± s.e.m. Statistical analysis was performed using two-tailed Student’s unpaired t-test (a, b, i, j, m, n, p-s), two-way ANOVA with multiple comparisons corrected using Šidák’s multiple comparison test (c, o), or one-way ANOVA with Tukey’s multiple-comparison test (e, f).

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