Fig. 3.

TAZ, but not YAP, is required for GPCR-mediated activation of the Hippo pathway and for thrombin-mediated migration and invasion. (A–C) MDA-MB-231 HA–ARRDC3 pSLIK cells were transfected with the indicated siRNAs and stimulated with 100 nM lysophosphatidic acid (LPA, A), 1 µM SLIGKV peptide agonist (B) or 100 nM sphingosine-1-phosphate (S1P, C). Samples were immunoblotted (IB) with antibodies against the indicated proteins. Results are represented as the fold-increase in CTGF and ANKRD1 expression relative to 0 h non-specific siRNA (NS)-transfected control. Data are mean±s.d., n=3. Statistical significance was determined by two-way ANOVA with Tukey′s post hoc test. (D,E) MDA-MB-231 HA–ARRDC3 pSLIK cells were transfected with the indicated siRNAs and incubated in transwells, with or without 100 pM α-thrombin (α-Th) or 0.5% FBS as control for migration assay (D), or with or without 1 pM α-thrombin for the invasion assay (E). Images are representative of three independent experiments. Scale bars: 20 μm. Results were quantified and are represented as the fold change over untreated control cells. Data are mean±s.d., n=3. Statistical significance was determined by one-way ANOVA (with Tukey′s post hoc test) for migration assay and unpaired t-test for invasion assay. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.