Figure 4.

YAP target genes are downregulated when the Hippo/YAP signaling pathway is activated by ectopically overexpressed N3ICD. (a and b) RT-PCR confirms that ectopically overexpressed N3ICD-induced upregulation of Kibra expression is epithelial cell specific, as this phenomenon does not occur in the U87 glioblastoma cell line. (c and d) Western blotting and immunofluorescent staining confirm that non-phosphorylated YAP is excluded from the nucleus (the white arrows show non-phosphorylated YAP in the nucleus by western blot) and then is phosphorylated and degraded in MDA-MB-231 cells ectopically overexpressing N3ICD as compared with control cells. (e) RT-PCR confirms that YAP target genes are downregulated once the Hippo/YAP signaling pathway is activated by ectopically overexpressing N3ICD. Data are presented as the mean±s.d. of three experiments. *P<0.05, **P<0.01 and ***P<0.001 (Student's t test) as compared with control cells.