Fig. 4: Inhibition of NETosis attenuates dissection progression in mice with AD.

A Schematic overview of experimental design. Each mouse received intraperitoneal injection of saline, Cl-amidine (10 mg/kg), DNase I (5 mg/kg) or both drugs at a fixed time once a day during 4–7 weeks of age until death or the end point of the experiment. B The incidence of AD and aortic rupture in each group (n = 12). Comparisons by Fisher’s exact test indicating aortic rupture rate difference among groups (p = 0.0137; p = 0.4783; p = 0.0137; p = 0.4783). C Mice death due to aortic rupture of each indicated group (n = 12) (p = 0.0378 for Cl-amidine vs BAPN; p = 0.0424 for Combo vs BAPN). D Representative vascular ultrasound images of aorta in each group (n = 12). Scale bar = 1 mm. E Representative macrographs of aortas in each group. Scale bar = 20 mm. F The average of max diameter (F) (p = 0.0018 for DNase I vs Ctrl), aortic wall thickness of aortas (G) (p < 0.0006 for BAPN vs Ctrl), and media thickness (H) in each group (n = 12). I Elastin degradation grading evaluation of aorta in each group (n = 12) (p = 0.0337 for Cl-amidine vs BAPN; p = 0.0436 for Combo vs BAPN). J Representative immunohistochemistry images showing aortic dilation, false lumen formation, and elastin degradation within aortas in each group. Arrow indicating broken elastin. (K) Representative immunofluorescence images showing NETs formation within aortas of mice, determined by CitH3 (citrullinated histone H3, green) and neutrophil (Ly6G, red) staining. Scale bar = 100 μm. Values are expressed as means ± SD. A two-sided statistical significance was set at *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by Mann–Whitney U test or Fisher’s exact test. Log-rank test was used for the analysis of survival rate.