Figure 7.

Wnt signaling regulates the accumulation of F-actin and nascent smooth muscle cells in the sub-epithelial mesenchyme to influence epithelial morphology

(A) Confocal sections of the left lobe of lungs isolated from E11.5 SMA-RFP embryos, immunostained for RFP, and labeled with phalloidin (F-actin) after treatment with DMSO or LiCl for 24 h.

(B) Quantification of F-actin intensity profiles emanating from the epithelium (n = 3–6).

(C) Quantification of RFP intensity profiles emanating from the epithelium of lungs isolated from SMA-RFP embryos (n = 4–5).

(D) E12.5 control and Tbx4-rtTA;tet-O-Cre;Ctnnb1^fl/fl lungs immunostained for F-actin.

(E) Quantification of F-actin intensity profiles emanating from the epithelium (n = 5).

(F) Quantification of Tbx3 intensity profiles emanating from the epithelium (n = 3).

(G) Confocal sections of the left lobe of lungs isolated from E11.5 CD1 embryos and immunostained for E-cadherin (Ecad) and αSMA after treatment with DMSO or LiCl for 24 hrs.

(H and I) Width and length of branch L.L2 after 24 h of culture (n = 13–16).

(J) Quantification of smooth muscle coverage of the left lobe after 24 h of culture (n = 6–11).

(K) E12.5 control and Tbx4-rtTA;tet-O-Cre;Ctnnb1^fl/fl lungs immunostained for Ecad and αSMA.

(L and M) Width and length of branch L.L2 after 24 h of culture (n = 7–8).

(N) Quantification of smooth muscle coverage of the left lobe in control and Tbx4-rtTA;tet-O-Cre;Ctnnb1^fl/fl lungs (n = 3). Data in bar graphs were plotted with error bars representing SD and were compared using t test. Intensity profiles were plotted as mean ± SEM and compared using two-way ANOVA. ∗ indicates p < 0.05, ∗∗ p < 0.001 and ∗∗∗ p < 0.0001.