Figure 6.

CXCL1 and PAD4 Are Required for the Adhesion of Bacteria to the Omentum in the Absence of Fluid Flow

(A) Representative confocal images of wholemount staining of omenta, isolated from either naive animals or 18 h after Zymosan i.p injection, in combination with the injection of either anti-CXCL1 or isotype control antibodies, and cultured with fluorescently labeled E. coli bio-particles for 10 min. Number of E. coli bioparticles found per cluster section and per μm² of cluster were graphed. MPO (white), E. coli (green), DAPI (red), Ly6G (blue). Data for cluster quantification pooled from two independent experiments with n ≥ 18 imaged clusters from n = 10–11 mice per group. Error bars show SEM.

(B) Mice were left naive (white circles) or injected i.p. with Zymosan in combination with the PAD4 inhibitor GSK484 (gray circles) or vehicle (blue circles). The omenta were collected 18 h post-injection and incubated in vitro for 5 min with mCherry⁺E. coli. Representative confocal images of wholemount staining of omentum. E. coli (green), DAPI (magenta). Mean gray value of E. coli in omFALCs and percentage area covered by E. coli.

Data for cluster quantification pooled from two independent experiments with n ≥ 10 imaged clusters from n = 7–8 mice per group. Error bars show SEM. Kruskal Wallis test with Dunn’s multiple comparisons test were applied after assessing normality using D’Agostino and Pearson normality tests, ns = non-significant, ^∗∗ = p < 0.01, ^∗∗∗p = < 0.001, ^∗∗∗∗p = < 0.0001. Scale bars, 50 μm.