Immune response in the tumor site
(A) Schematic diagram of IFN production after treatment. (B) Cytokine levels (IFN-β, IFN-γ, TNF-α, IL-12, IL-10, and TGF-β1) in excised tumors after different treated mice by ELISA analysis. G1, PBS; G2, siAdar1-LNP@m; G3, siNC-LNP@mPD1; G4, siAdar1-LNP@mPD1 (mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; n = 3). (C–F) Typical FCM plots and quantification results of CD3+CD8+ T cells (C), CD3+CD4+ T cells (D), Tregs (E), and MDSCs (F) in tumors of four different treatment groups (mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; n = 3). (G) Multiple immunofluorescence images displayed the infiltrated CD4+ T cells (red) and CD8+ T cells (green) within tumors tissue. Scale bar, 100 μm. An enlarged image of the upper panel is shown in the lower panel. Scale bar, 25 μm. (H) Volcano plot of disregulated genes in tumor tissues with siAdar1-LNP@mPD1 treatment versus PBS treatment. (I) Heatmap showing disregulated genes in mice tumors after siAdar1-LNP@mPD1 and PBS treatments (fold change ≥ ± 10, false discovery rate < 0.05; n = 3). (J) KEGG enrichment analysis of the disregulated genes between siAdar1-LNP@mPD1 and PBS treatment.