Fig. 9.

FZD₄-induced and Gα_12/13-mediated p115-RHOGEF-GFP membrane recruitment depends on activation of the heterotrimeric Gα_12/13 proteins. HEK293 cells were transfected with FZD₄-Cerulean, Gα₁₂-mCherry (A) or Gα₁₃-mCherry (B), and p115-RHOGEF-GFP either without or with the isolated AU1-tagged RGS domain of p115-RHOGEF-RGS. Cells presenting membranous p115-RHOGEF-GFP localization were counted, and data from four independent experiments were summarized in the bar graph. More than 50 cells were counted for each condition in each independent experiment. Values give the mean ± S.E.M. (C) Cells used for quantification of p115-RHOGEF-GFP recruitment were lysed and immunoblotted for vinculin (loading control) and anti-AU1 to detect the tagged-RGS domain of p115-RHOGEF.