Figure 4. Fusobacterium nucleatum aggravates colorectal cancer liver metastasis by restructuring the liver immune microenvironment. (A) Myeloid-derived suppressor cells (MDSCs; CD45+CD11b+Gr-1+) in the liver and spleen were determined by flow cytometry. (B) NK (CD45+CD3-CD49b+) and NKT cells (CD45+CD3+CD49b+) were analyzed by flow cytometry. (C, D) CD3+T cells (C), CD3+CD4+T cells and CD3+CD8+T cells (D) in the liver and spleen were assessed by flow cytometry and analyzed by gating on lymphocytes and CD3+ cells. (E) Tumor-associated macrophages (CD45+CD11b+F4/80+) in the liver and spleen were determined by flow cytometry. (F) Treg cells (CD4+Foxp3+) and Th17 cells (IL17+CD4+) were evaluated by flow cytometry and analyzed by gating on CD4+T cells. Data are presented as mean ± standard deviation (n=5 per group). *P < 0.05, **P < 0.01, and ***P < 0.001; unpaired Student’s t-test.