Skip to main content
. 2024 Jul 26;10(30):eadn5405. doi: 10.1126/sciadv.adn5405

Fig. 4. CDH lungs have an inflammatory phenotype with high macrophage density that is reduced to normal levels by AFSC-EV administration.

Fig. 4.

(A) UMAP of snRNA-seq data split by condition. (B) Violin plots of macrophage and inflammatory marker gene expression across cell types, as measured by snRNA-seq. (C) Representative immunofluorescence images of pan-macrophage marker CD68 in rat fetal lungs from all three conditions, quantified as fluorescence intensity of CD68 per field. Scale bars, 50 μm. Control+saline (n = 8), CDH+saline (n = 6), and CDH+AFSC-EV (n = 8). Groups were compared using Kruskal-Wallis (post hoc Dunn’s nonparametric comparison) for (C), according to Shapiro-Wilk normality test. ***P < 0.001; ****P < 0.0001. (D) Flow cytometry analysis of dissociated lung cells stained for CD68 (red) versus unstained (black) and (E) costained with ADGRE-1 and CD43 (panels are representative of n ≥ 3 pups per group; data file S4). *P < 0.05; **P < 0.01. (F) Representative histology images (hematoxylin and eosin) of fetal lungs from Control+saline, CDH+saline, and CDH+GW2580 fetuses. Scale bars, 50 μm. (G) Differences in number of alveoli (RAC) in Control+saline (n = 7), CDH+saline (n = 8), and CDH+GW2580 (n = 8), quantified in at least seven fields per fetal lung. (H) Gene expression changes in inflammatory markers Tnfα and Lcn2 in RAW264.7 cells stimulated with LPS, relative to Actb housekeeping gene.