Figure 4.

CitH3⁺ DNA Coats Neutrophil Aggregates on omFALCs during Peritonitis

(A) Representative confocal images of wholemount staining of omentum from naive and 6 h post-Zymosan mice DAPI (magenta), CitH3 (green), and Ly6G (blue). Scale bar, 50 μm.

(B) Quantification of the number of CitH3 positive nuclei per mm at the times indicated post-Zymosan injection. Data for cluster quantification pooled from two independent experiments with n ≥ 24 imaged clusters from n = 8 mice per group. Box and whiskers showing min to max value.

(C) 3D reconstruction of omFALC at 6 h post-Zymosan injection showing the surface of a cluster (first row) and a clipped view inside the cluster (second row). Ly6G (blue), CitH3 (green), DAPI (magenta). Scale bar, 100 μm.

(D) WT or Elane^−/− mice were left naive (WT white, Elane^−/− gray) or injected i.p. with Zymosan (WT blue, Elane^−/− gray). Omenta were collected 18 h post-injection. The number of CitH3⁺ cells per mm and volume of each cluster were assessed by wholemount staining and confocal analysis of omenta from naive and treated mice. Data for cluster quantification pooled from two independent experiments with n ≥ 14 (CitH3⁺ cells) or n ≥ 20 (cluster size) imaged clusters from n = 4 mice per group.

(E) Representative confocal images of omFALCs at 6 h following Zymosan injection showing MPO (green), Ly6G (blue), CitH3 (white), and DAPI (magenta). Staining representative of n ≥ 24 clusters from n ≥ 8 mice in two independent experiments. Scale Bar, 50 μm.

Student’s t test, Kruskal Wallis test with Dunn’s multiple comparisons test, or Mann Whitney test was applied after assessing normality using D’Agostino and Pearson normality test, ns = non-significant, ^∗∗∗∗p = < 0.0001.