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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Oncogene. 2015 Mar 23;34(50):6040–6054. doi: 10.1038/onc.2015.52

Fig. 6. YAP regulates expression of EGF-like ligands and ERBB receptors.

Fig. 6

a) Determine the mRNA expression of NRG1, NRG2, HBEGF, EGFR and ERBB3 in HOSE-MXIV, HOSE-YAP and HOSE-YAPS127A cells (left panel), and in TOV21G-MXIV, TOV21G-YAP & TOV21G-YAPS127A cells (right panel) using RT-PCR. b) Determine protein levels of YAP, phosphorylated YAP [p-YAP (S127)], EGFR, and ERBB3 in HOSE-MXIV, HOSE-YAP and HOSE-YAPS127A cells (left panel), and in TOV21G-MXIV, TOV21G-YAP & TOV21G-YAP S127A cells (right panel). c) Determine the expression of EGFR (left, green) and ERBB3 (right, green) in xenograft tumors derived from the TOV21G-MXIV, TOV21G-YAP & TOV21G-YAPS127A cell lines using Fluorescent immunohistochemistry. Actin was stained with rhodamine-phalloidin (red). Nuclei were stained with DAPI (blue). Scale bar: 20μm. d) mRNA levels of ERBB1, ERBB3, HBEGF and NRGs in the tumor xenografts derived from TOV21G-MXIV, TOV21G-YAP, and TOV21G-YAPS127A cells. e) Top panel: Western blot analysis to detect the expression of YAP, EGFR and ERBB3 protein in TOV21G cells with or without YAP knockdown using YAP siRNA (si-YAP). siGLO (a non-target siRNA) was used as a negative control; Lower panel: mRNA levels of NRG1, NRG2 and HBEGF before and after Knockdown of YAP with YAP siRNA (si-YAP). f & g) Concentrations of HBEGF and NRG1β1 in the medium of HOSE-MXIV, HOSE-YAP & HOSE-YAP S127A cells. h & i) Concentrations of HBEGF and NRG1β1 in the medium of TOV21G cells transfected with non-targeting control siRNA (siGLO) or YAP siRNA (siYAP). All experiments were repeated at least three times and the representative images were presented. Bars with different letters are significantly different from each other (p < 0.05).