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Fig. 6. Dia1 localizes to and strengthens the adherens junctions in a Rho-dependent manner. (A-C) GFP-tagged full-length mDia1 (GFP-mDia1) is distributed diffusely all over the transfected cells (A). The distributions of F-actin (B) and 
-catenin (C) in cells expressing GFP-mDia1 are not affected. (D-F) Co-transfection of GFP-mDia1 together with the constitutively active mutant of RhoA (RhoA-V14) leads to a significant enrichment of GFP-mDia1 at cell-cell junctions (D) and formation of numerous filopodia-like projections, with GFP-mDia1 localized to their tips (D, inset). It also promotes formation of prominent phalloidin-stained actin cables (E) and a significant accumulation of -catenin at the interface between transfected cells (F). (G-I) Transfection of RhoA-V14 strongly enhances the localization of endogenous Dia1 to cell-cell junctions, as revealed by staining with antibodies against Dia1 (G), promotes formation of stress fibers (H) and induces some increase in cell-cell junctions marked by staining for -catenin (I). Arrows indicate the contacts between cells. (J) Assessment of the Rho-mediated localization of GFP-mDia1 to cell-cell contacts. The fluorescence intensity of GFP-mDia1 was measured by line-scan analysis at cell-cell interfaces. A localization index was calculated as described in Materials and Methods. Means±s.d. are shown. (K) The fluorescence intensities of actin and -catenin at cell-cell contacts were measured as explained in the legend to Fig. 3. Bars represent the average fluorescence intensity expressed in arbitrary units. Error bars represent s.d. values. Asterisks indicate the values that differ significantly from those of corresponding controls (according to both Kolmogorov-Smirnov (KS) and Student's t-test, P<0.001). Quantifications were done for three independent experiments, including measurements of 30 pair of cells for each experiment.
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