Figure 5.

Inhibition of PAD4 Prevents Neutrophil Aggregation and Capture of Zymosan Particles within omFALCs While Increasing the Retention of Zymosan in the Peritoneal Cavity and Spread to the Spleen

Mice were left naive (white) or injected i.p. with Fluo-Zym in combination with the PAD4 inhibitor GSK484 (red) or vehicle (blue) and the omentum. PECs and spleen were analyzed 6 h post injection.

(A and B) Representative low (A, scale bar 1 mm) and high (B, scale bar 50 μm) magnification confocal images of wholemount immunofluorescence staining of omentum.

(C–E) Number of Fluo-Zym particles per mm² of omFALCs (C), mean gray value for CitH3 and Ly6G staining (D), volume of omFALCs (E).

(F) Total number of PEC neutrophils.

(G) Representative histogram showing fluorescence intensity of Fluo-Zym in all PECs and number of Fluo-Zym⁺ PECs.

(H) Number of Fluo-Zym⁺ neutrophils and MFI of Fluo-Zym within Fluo-Zym⁺ neutrophils.

(I) Representative histogram showing fluorescence intensity of Fluo-Zym in spleen and percentage and number of Fluo-Zym⁺ events per spleen.

Data for cluster quantification pooled from two independent experiments with n ≥ 30 (C–D) and n ≥ 60 (E) imaged clusters from n = 7–8 mice per group. Data pooled from two independent experiments with n = 7–8 mice per group. 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 < 0.01, ^∗∗∗p = < 0.001, ^∗∗∗∗p = < 0.0001.