Fig. 5.

DVL is required for cytoplasmic trafficking of YAP induced by E-cadherin or α-catenin. a The wt and DVL-TKO 293 T-REx cells were transfected with flag-tagged YAP (50 ng) in combination with vector control (1 μg) or E-cadherin (1 μg) or α-catenin (1 μg), and YAP localization was determined by confocal microscopy. Inset, DAPI nuclear stain; Scale bar, 5 μm. b The wt and DVL-TKO 293 cells were transfected with vector control (−) or E-cadherin (E-cad) or α-catenin (α-cat), and endogenous YAP abundance in nuclear fraction and whole-cell lysates (WCL) was determined by immunoblot analysis. Relative nuclear YAP abundance compared to control was measured by ImageJ. c The MCF-10A cells were cultured under confluent contact inhibition and intracellular YAP localization was monitored by confocal microscopy. The cells were incubated with a mouse IgG (HECD-) or a neutralizing monoclonal antibody (HECD, 10 μg/ml) that disrupts homophilic binding of E-cadherin for 16 h. Immunofluorescence images showing YAP staining of MCF-10A cells having tetracycline-inducible DVL3 in the absence (Dox-) or presence (Dox+) of doxycycline. Upper inset, HECD antibody detected by fluorescent-conjugated secondary antibody; lower Inset, DAPI nuclear stain; Scale bar, 5 μm. Unprocessed original scans of blots are shown in Supplementary Fig. 10