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. Author manuscript; available in PMC: 2016 Jan 21.
Published in final edited form as: Oncogene. 2014 Sep 8;34(27):3536–3546. doi: 10.1038/onc.2014.281

Figure 3.

Figure 3

vGPCR activates YAP/TAZ. (a) vGPCR expression increases YAP and TAZ protein levels. MCF10A, HEK293A, and SVEC cells stably expressing either the vector control or HA-vGPCR were serum-starved for 12 hours before immunoblotting analysis. (b) vGPCR stabilizes YAP/TAZ. HEK293A cells stably expressing vGPCR were treated with cycloheximide for the indicated time (hours: hr). (c) vGPCR induces YAP/TAZ nuclear localization. HEK293A cells overexpressing either control (CTL) or vGPCR were serum-starved for 12 hours. YAP/TAZ subcellular localization were determined by immunofluorescence staining for YAP/TAZ (red), TAZ (green), along with DAPI for DNA (blue). (d) vGPCR activates a YAP/TAZ reporter. Control (CTL), vGPCR, or positive control LPAR plasmids were co-transfected with a 5 X UAS-luciferase reporter, Renilla and Gal4-TEAD4 into HEK293A cell. 24 hours after transfection, cells were serum starved for 12 hours and luciferase activity was measured and quantified by normalization to the co-transfected Renilla. (e) vGPCR induces expression of YAP/TAZ target genes. mRNA levels of indicated YAP/TAZ target genes was measured in stably expressing control (CTL) and vGPCR cells following 12 hour starvation.