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. 2020 Apr 14;52(4):700–715.e6. doi: 10.1016/j.immuni.2020.03.011

Figure 3.

Figure 3

CXCL1 Is Required for the Recruitment of Neutrophils into omFALCs

(A) Number of neutrophils in digested omentum as assessed by flow-cytometric analysis (Figure S5A) of naive (white bar) and at the indicated time points following i.p. injection of Zymosan-A (blue bars). Data pooled from two independent experiments with n = 5–11 mice per group.

(B) Representative confocal imaging and 3D reconstruction of omFALCs from naive and 6 h post-Zymosan mice, DAPI (magenta) and Ly6G (white). Scale bar, 200 μm.

(C) Quantification of the mean gray value of Ly6G and MPO stained as in Figure S5B of omenta from naive (white bar) and at the indicated time points following i.p. injection of Zymosan (blue bars). Data for cluster quantification pooled from two independent experiments with n ≥ 24 imaged clusters from n = 8 mice per group.

(D) Quantification of the volume of omFALCs at the indicated times following i.p. injection of Zymosan-A (blue). Data pooled from two independent experiments with n ≥ 32 imaged clusters from n = 6 mice per group.

(E) Representative confocal images of omentum, the peritoneal surface of the diaphragm, and parietal wall from naive and 6 h post-Zymosan mice, DAPI (magenta) and Ly6G (white) and quantification of the mean gray value of Ly6G for omFALCs, parietal wall, diaphragm from naive (white) and 6 h post-Zymosan mice (blue). Data for cluster quantification pooled from two independent experiments with n ≥ 24 imaged clusters from n = 6 mice per group. Scale bar, 50 μm.

(F) Representative density plots showing proportion of neutrophils found in the peritoneal exudate cells (PECs) and omentum 18 h post Zymosan i.p injection, in combination with the injection of anti-CXCL1 or isotype control antibodies after 2 h and number of neutrophils found in PECs and per g of omentum in naive and treated mice. Data pooled from two independent experiments with n = 5–11 mice per group.

Error bars show SEM. Box and whiskers showing min to max value. Kruskal Wallis test with Dunn’s multiple comparisons test or ANOVA with Sidak’s multiple comparisons test was applied after assessing normality using D’Agostino and Pearson normality tests, ns = non-significant, ∗ = p < 0.05, ∗∗ = p < 0.01, ∗∗∗p = < 0.001, ∗∗∗∗p = < 0.0001.