Figure 1. Activation of YAP by mutant Gq/11 in UM cell lines.

(A) Effects of UM-associated mutant Gq/11 on YAP/TAZ activity. HEK293A cells were transfected with different Gq/11 plasmids together with FLAG-YAP, and following 12 hr serum starvation, cells were harvested and YAP/TAZ activation status was analyzed by immunoblotting using the indicated antibodies. YAP phosphorylation was assessed using gels containing phos-tag, which slows down migration of phosphorylated YAP during electrophoresis. Endogenous TAZ protein levels were determined with a TAZ specific antibody. For the Gq plasmids, two concentrations of plasmids were used in transfection and the expression levels of transfected Gq were determined by immunoblotting. (B) A table summarizing Gq/11 or BRAF mutation status, YAP phosphorylation, YAP localization, and responses to serum of multiple UM cells. Nuc, Nucleus; Cyto, cytoplasm; wt, wild type; the number of “+” indicate the strength; *, information from Griewank et al., 2012. (C) YAP phosphorylation and response to serum in representative UM cell lines. UM cells were cultured with or without 10% FBS for 16 hr. (D) YAP localization and response to serum in representative UM cell lines. Cells were maintained in the presence or absence of 10% FBS for 16 hr, and after fixation, YAP localization was determined by immunostaining. The green and blue colors represent YAP and DNA staining, respectively. Scale bars represent 5 µm.

See also Figure S1.