Fig. 2. Major hepatectomy increases hepatocellular EGFR levels and contributes to YAP1 activation.
(A) GSEA of RNA-seq data from mouse livers 3 days after hepatectomy. (B) Heatmap of gene expressions of YAP1 conserved signature in RNA-seq data from mouse liver in sham, 30% PHx, and 70% PHx groups 3 days after surgeries. (C and D) IHC and WB analyses of YAP1S127 and nuclear YAP1 in the remaining livers. (E) qRT-PCR analysis of YAP1 downstream genes Ccn1 and Ccn2 levels in the remnant liver. (F) Schematic of the CRLM mouse model with/without YAP1 overexpression in hepatocytes generated via AAV8. (G and H) Liver–to–body weight ratios and tumor-to-liver volume ratios of the negative control (NC) and overexpression (OE) groups. (I) Schematic, images, IF, and hematoxylin and eosin (H&E) staining of livers from the NC or OE groups. (J) Survival curve of the CRLM model mice in the NC and OE groups. (K) Schematic of the CRLM mouse model with/without hepatocellular YAP1 overexpression and verteporfin administration. (L and M) Liver–to–body weight ratios and tumor-to-liver volume ratios of the mice in the NC, YAP1 OE, and YAP1 OE with verteporfin groups. (N) Schematic, images, MRI scans, and IF and H&E staining of livers and lungs from CRLM model mice in the above three groups. (O) Survival curve of the CRLM model mice in the above three groups. (P and Q) WB and IHC analyses of EGFR levels in the remnant liver tissue in sham, 30% PHx, and 70% PHx groups. (R to T) WB and IF analyses of YAP1 levels in AML12 cells after the addition of EGF and canertinib. (T) WB analysis of EGFR and YAP1 levels after major hepatectomy and the addition of canertinib. (U) CoIP analysis showing the combination of the YAP1 and EGFR proteins. IgG, immunoglobulin G.