Fig. 7: In vivo prevention of primary colon tumor occurrence and metastasis.

a Schematic illustration of the pre-treatments with the given agents (once-daily oral gavage for 10 days), biochemical and immunological analyses on blood, spleen, and MLN samples, initial inoculation and rechallenge of colon cancer cells, and macrophage or CD8⁺ T cell (Mac/CD8 T) depletion. Created in BioRender. Cao, P. (2025) https://BioRender.com/l8qpogu. b Bioluminescence imaging of the colon tumors in the mice receiving different treatments at the given time points after initial orthotopic tumor inoculation (n = 5). c Bioluminescence imaging and representative images of orthotopic tumor development in mice after the rechallenge of colon cancer cells. The dotted circles mark the location of the tumor. Scale bar, 1 cm. d Schematic illustration of the prevention in transgenic APC^min/+ mice receiving the given treatments for 9 weeks. Created in BioRender. Cao, P. (2025) https://BioRender.com/l8qpogu. e, f Representative images and the corresponding quantification of tumors in the intestine of APC^min/+ mice after different treatments (n = 5). Red triangular arrows indicate the tumor nodules. Scale bar, 1 cm. Representative images of H&E staining (g) and immunohistochemistry staining of Ki67 (h) of the intestinal segments. Blue triangular arrows indicate the tumor nodules. Scale bar, 2.5 mm. i, j Quantification of the ratio of M1 (CD11b⁺F4/80⁺CD86⁺) to M2 (CD11b⁺F4/80⁺CD206⁺) macrophages, and the percentages of mature DCs (CD11c⁺CD86⁺) in total CD45⁺ cells within the spleen from the mice receiving the given treatments for 10 days (n = 5). k, l Quantification of the percentages of activated CD4⁺ and CD8⁺ T cells (IFN-γ⁺CD4⁺ and IFN-γ⁺CD8⁺) in total CD3⁺ cells within the MLNs from the mice receiving the given treatments once daily for 10 days (n = 5). m Schematic illustration of the establishment of the liver metastasis model by injecting MC38 cells into the spleen of mice, once-daily oral gavage of the given agents, and macrophage or CD8⁺ T cell (Mac/CD8 T) depletion. Created in BioRender. Cao, P. (2025) https://BioRender.com/l8qpogu. n Representative images of the liver and spleen tissues isolated from the mice 14 days after tumor inoculation. Blue triangular arrows indicate the liver tumor nodules. o The weight of livers and spleens isolated from the mice in each group 14 days after tumor implantation (n = 5). p The quantification of liver tumor nodules in mice in each group 14 days after tumor inoculation (n = 5). The “n” represents the number of biologically independent samples. Data are shown as mean ± SD; p-value (compared to PBS group); one-way ANOVA (one-tailed), Tukey’s multiple comparisons. Source data are provided as a Source Data file.