Figure 3.
Wnt signaling regulates cell identity in the embryonic pulmonary mesenchyme.
(A) Bubble plot showing the enrichment percentage and adjusted p value of relevant GO terms identified based on genes upregulated in each mesenchymal cluster.
(B) Heatmap showing the expression of Wnt-associated genes upregulated in either mesenchymal cluster. Activators and targets are colored in blue, inhibitors are colored in red.
(C) Heatmap showing the expression of Wnt ligands, secreted inhibitors, and receptors detected in either mesenchymal cluster and in clusters containing cells from the mesothelium (meso), vascular endothelium (ve), epithelium (ep), and smooth muscle (sm).
(D–G) Confocal sections and quantification of Lef1 and Foxp1 intensity profiles around branch L.L2 in lungs isolated at E11.5 from CD1 embryos and immunostained for Lef1 or Foxp1 after treatment with either DMSO, LiCl (10 mM), or IWR1 (100 μM) for 24 h (n = 2–6). Yellow dashed lines indicate the border of the epithelium. Schematics show lines and direction along which intensity profiles were measured. Mean and SEM are plotted, and curves were compared using two-way ANOVA.
(H–M) E12.5 control and Tbx4-rtTA;tet-O-Cre;Ctnnb1fl/fl lungs immunostained for Lef1 or Foxp1 and quantification of Lef1 and Foxp1 intensity profiles (n = 3). Low-magnification z-projections (H and I) and high-magnification confocal slices (J and K) are shown. Scale bars show 50 μm. ∗ indicates p<0.05, ∗∗ indicates p<0.001, and ∗∗∗ indicates p<0.0001