Fig. 5

Fn-induced NETs facilitate malignant growth and metastasis in vivo. A The schematic diagram of Fn inoculation by gavage with and without DNase I treatment based on AOM/DSS mice model. B Representative images of the whole colon tissues (left panel) and the tumor nodules in colon tissues as indicated by the red arrows (right panel). C Colon length from all groups of mice was measured on week 12. Data are shown as means ± SD. D The colonic tumor nodules from all groups of mice were counted on week 12. Data are shown as the average tumor number ± SD. E Representative images of H&E staining, FISH for Fn, IF staining for CitH3 and IHC staining for Ly6G, Ki67, CD31, VEGF, MMP2 and MMP9 in representative colon tumor sections from AOM/DSS mice treated with or without Fn and DNase I. Blue scale bars: 100 µm. White scale bars and black scale bars: 50 µm. F The schematic diagram of CRC metastasis mice injected with HCT116 cells, HCT116 cells + Fn or HCT116 cells + Fn + DNase I. G Representative images of morphological features of macroscopic lungs of nude mice and the H&E staining of lung metastasis. Black scale bars, 50 µm; Red scale bars, 100 µm; Green scale bars, 200 µm; Blue scale bars, 400 µm. H Statistical analysis of metastasis focuses in lungs from individual mouse. I ELISA analysis for the serum levels of MPO-DNA in mice. J The schematic diagram of purified NETs treatment based on AOM/DSS mice model. K Representative images of the whole colon tissues (left panel) and the tumor nodules in colon tissues as indicated by the red arrows (right panel). L Colon length from all groups of mice (n = 4 / each group) was measured on week 12. Data are shown as means ± SD. M The colonic tumor nodules from all groups of mice (n = 4 / each group) were counted on week 12. Data are shown as the average tumor number ± SD. N Representative images of H&E staining, IF staining for CitH3 and IHC staining for Ly6G, Ki67, CD31, VEGF, MMP2 and MMP9 in representative colon tumor sections from AOM/DSS mice treated with or without purified NETs. Blue scale bars: 100 µm. White scale bars and black scale bars: 50 µm. O The schematic diagram of CRC metastasis mice injected with HCT116 cells or NETs-treated HCT116 cells. P Representative images of morphological features of macroscopic lungs of nude mice and the H&E staining of lung metastasis. Black scale bars, 50 µm; Red scale bars, 100 µm; Green scale bars, 200 µm; Blue scale bars, 400 µm. Q Statistical analysis of metastasis focuses in lungs from all groups of mice (n = 5 / each group). *p < 0.05, **p < 0.01, ***p < 0.001