Fig. 6

The effect of NETs on the proliferation and migration of HUVECs. a The proliferation ability was detected using the CCK-8 method under different NET concentrations: 0, 0.2, 0.4, and 0.8 μg/mL. b EdU labeling was used to detect the effect of NETs on the proliferation of HUVECs and the inhibitory effect of DNase-1. Magnification, × 20; scale bars: 50 μm; red, EdU; blue, Hoechst. c The HUVEC proliferation rate under EdU labeling; all values are presented as means ± SDs. ****p < 0.0001. d, e Western blotting to detect changes in VE-cadherin and ZO-1 expression under stimulation with different NET concentrations: 0, 0.2, 0.4, and 0.8 μg/mL. f, g Image J software was used to calculate the gray values of VE-cadherin and ZO-1 under different NET concentrations; *p < 0.05, ****p < 0.0001. h The effect of NETs on the expression of VE-cadherin and ZO-1 and the inhibitory effect of DNase-1 as observed by immunofluorescence staining. Magnification, × 20; scale bars: 50 μm; red, F-actin; green, VE-cadherin/ZO-1; blue, DAPI. i, j The average fluorescence intensities of VE-cadherin and ZO-1 were calculated using Image J software. All values are presented as means ± SDs. *p < 0.05; **p < 0.01; ***p < 0.001. k The effect of NETs on the migration of HUVECs and the inhibition of DNase-1 as evaluated using the scratch test. Magnification, × 20; scale bars: 50 μm. l The percentage of cell migration area was calculated using Image J software. All values are presented as means ± SDs. **p < 0.01; ****p < 0.0001. NET, neutrophil extracellular trap; DNase-1, deoxyribonuclease I; SD, standard deviation