AIM2-silenced DC vaccine improves the efficacy of ACT against melanoma. (A) Immunoblotting for AIM2 and vinculin in the lysates of mock-, control siRNA–, or Aim2 siRNA– (-1 or -2) transfected WT BMDCs 48 h after transfection. (B) Quantitative RT-PCR analysis of the Aim2 mRNA expression in mock-, control siRNA–, or Aim2 siRNA–transfected WT BMDCs 2, 10, and 22 d after transfection (n = 6). (C–E) B16F10 mice were treated with ACT + control siRNA– or Aim2 siRNA–transfected WT DC-gp100. On day 20 after PMELs transfer, tissues were harvested. (C) Therapy regimen scheme. (D) Tumor growth over time (left; n = 9). Sample photo of B16F10 tumor on day 20 after PMELs transfer (right). Scale bar, 10 mm. (E) Flow cytometry analysis of the numbers of PMELs, CD8+, and CD4+ T cells among 104 live singlet cells, percentage of FoxP3+ cells in CD4+ T cells, and PMEL/T reg cell ratio in the tumor (n = 9). Data are shown as mean ± SEM and are representative of three independent experiments (A) or are pooled from two independent experiments (B, D, and E). *, P < 0.05; **, P < 0.01; ***, P < 0.001; two-way ANOVA with Tukey’s multiple-comparisons test (D) or one-way ANOVA with Dunnett’s multiple-comparisons test (B and E).