FIGURE 1:

YAP and EDTB colocalize on endosomes. (A) MDCK cells grown on coverslips were labeled for endogenous EDTB, YAP, and EEA1. YAP colocalizes with EDTB (arrowheads) on intracellular puncta with a Pearson r of 0.31, whereas the Pearson r of YAP with EEA1 is 0.18. Scale bars, 10 μm (top), 2.5 μm (inset). (B–D) EDTB overexpression induces enrichment of YAP in the nucleus. MDCK cells expressing full-length EDTB (B, ET-FL) or control plasmid were grown on coverslips to confluence, and YAP localization was assessed using immunofluorescence. DAPI staining was used to determine total cell number. Representative images. (C) Quantification of B. ImageJ was used for quantification of nuclear enrichment of YAP protein. An identical threshold was set for all images, and YAP-positive nuclei were counted using the analyze particles function of ImageJ. Error bars, SEM of percentage of cells with nuclear enrichment of YAP; *p < 0.05. (D) GFP-CD and control cells grown to confluence and labeled with YAP. Expression of GFP-CD results in increased nuclear YAP (arrows).