2.7. Analysis of YAP activity

We used a luciferase based assay developed previously (Tian et al., 2010) to monitor YAP activity. We used two plasmids, one containing GAL4-TEAD construct, a gift from Dr. Kunliang Guan (Addgene plasmid #24640) and the other containing UAS-luciferase cassette, a gift from Dr. Liqun Luo (Addgene plasmid #24343) (Potter et al., 2010). To generate adenoviruses we cloned the GAL4-TEAD or the UAS-luciferase fragments into the pENTR11 shuttle vector and then transferred these to the adenovirus vectors pAd-DEST or pAd-CMV-DEST (Invitrogen) using the Gateway vector system (Invitrogen) producing the pAd-CMV-GAL4-TEAD and pAd-UAS-luciferase. Adenovirus was generated by transfecting the adenovirus plasmids into HEK293 cells.

To detect the YAP subcellular location, we used the GFP-YAP construct, a gift from Dr. Marius Sudol (Addgene plasmid #17843) (Basu et al., 2003). Adenovirus expressing the GFP-YAP construct was generated using the Gateway system as described above.