Figure 6.

YAP activation increases expression of AT1 and proximal epithelial cell signature genes

RNA-seq analyses were performed on EPCAM+ cells isolated from D14 YAP^active (N = 6) and controls (N = 4) mice.

(A) Volcano plot of the differentially expressed genes (p < .05, FC > 1.5) shows increase in AT1 markers Ager and Aqp5 as well as proximal lung airway markers Muc5b, Scgb1a1, Scgb3a1 and Scgb3a2.

(B) Genes upregulated in the YAP^active epithelium are associated with mitotic cell cycle and epithelial cell differentiation and development.

(C) Functional enrichment analyses of the subset of genes associated with epithelium development revealed increases in the expression of genes associated with Wnt signaling, epithelium proliferation, IPF and abnormal AT1 differentiation.

(D) Realtime qRT-PCR analyses of isolated YAP^active epithelial cells demonstrate increased Ager, Aqp5, Elf3 and Klf5. ∗Indicates p<0.05 as determined by Welch's t-test.

(E) Immunofluorescence analysis of NFIB (red), KLF5 (green), and SFTPC (white) shows presence of KLF5 in SFTPC+ cells including the SFTPC+ AT2 cell doublets identified in YAP^active mice.

(F) qRT-PCR analysis of proximal cell signature genes demonstrate increased conducting airway epithelial cell markers in YAP^active epithelial cells. ∗Indicates p<0.05 as determined by Welch's t-test.

(G) Immunofluorescence analysis of MUC5B (green), SFTPC (red), and SCGB1A1 (white), shows the presence of SCGB1A1+/SFTPC+ cells in YAP^active mice.

(H) RNAscope Fluorescent in-situ hybridization of Muc5b, Scgb1a1, and Sftpc in WT and YAP^active mouse lungs demonstrating the presence of Sftpc+/Muc5b+ cells in YAP^active lungs.

(I) Promoter analyses of the genes increased in the YAP^active epithelium show that transcription factors Klf5, Elf3, Nr2f6, and Mycn RNAs are increased and have enriched binding sites in YAP^active induced genes. See Table S1 for pathways down regulated in YAP^active mice and Figure S4 for immunofluorescence images.