Figure 2.

FALCs Are Covered by a Differentiated Monolayer of Mesothelial Cells

(A) 3D reconstruction of a large portion of the omentum obtained by imaging of wholemount staining of the omentum showing omFALCs with DAPI (white).

(B and C) Confocal imaging and 3D reconstruction of an omFALC showing a view of the surface of the cluster and a z section (along the dotted line) of the cluster (B) and of the surface of the omentum (C) with PDPN (red), CXCL13 (green), DAPI (blue), and IgM (yellow).

(D–F) Representative confocal imaging and 3D reconstruction of an omFALC showing a view of the surface of the cluster and a z section (dotted line) of the cluster with PDPN (red); ENPP2 (D, green), Cathepsin-C (E, CTSC, green), or ISG15 (F, green); DAPI (blue) and IgM (D, yellow); or CD45 (E and F, yellow). The surface of the cluster and the mesothelium are delimited by a hyphenated line.

(G) Violin plots of gene expression of inflammatory chemokines by cluster with highest log-normalized expression value labeled.

(H) Amounts of CCL2, CXCL10, and CXCL1 secreted into the supernatant of 2-h omentum explant culture per omentum and per ml after exposure to Zymosan-A for 2 h either in vivo (i.p. injection) or ex vivo. Data pooled from two independent experiments with n = 10 mice per group.

(I) Confocal imaging and 3D reconstruction of omFALC showing the surface of omFALCs (first column) and a clipped view inside the cluster (second column). Scale bars, 100 μm.

All staining representative of n ≥ 8 clusters from n ≥ 4 mice in at least two independent experiments. Error bars show SEM. 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 test, ns = non-significant, ^∗∗∗p = < 0.001.