Fig. 3

The effect of NETs on survival of CRC cells in vitro. A-E HUVEC tube formation assay using various CM from CRC cells treated with (Neu)-CM, (Neu + Fn)-CM, (Neu + Fn + Dnase I)-CM or with and without NETs for 4 h (A). Quantification analysis of the average branch number (B-E). F-G HUVEC tube formation assay treated with and without NETs for 4 h (F). Quantification analysis of the average branch number (G). H Western blot analysis of VEGF in CRC cells exposed to (Neu)-CM, (Neu + Fn)-CM, (Neu + Fn + DNase I)-CM or with and without NETs for 48 h. Protein levels of VEGF were quantified by using densitometry and normalized to β-actin and are shown as fold changes compared to the control (Right). Each bar displays the means ± SD of three independent experiments. I-J CCK8 assay for proliferation ability of CRC cells cultured with (Neu)-CM, (Neu + Fn)-CM, (Neu + Fn + DNase I)-CM, (Neu + PMA)-CM for 24, 48, and 72 h. K-L CCK8 assay for proliferation ability of CRC cells treated with or without NETs for 24, 48, and 72 h. M–O Apoptosis analysis for CRC cells treated with (Neu)-CM, (Neu + Fn)-CM, (Neu + Fn + DNase I)-CM for 48 h. Data are shown as means ± SD in three independent experiments (N–O). P-R Apoptosis analysis for CRC cells treated with or without NETs for 48 h. Data are shown as means ± SD in three independent experiments (Q-R). ns, not significant, White scale bars: 200 μm.*p < 0.05, **p < 0.01, ***p < 0.001