Fig. 5. Subcellular localization of p115RhoGEF in cells co-expressing constitutively active α₁₃ mutants.

HEK293 cells were transiently co-transfected with 0.5μg each expression plasmid for the indicated α₁₃ constructs and 0.1 μg DNA encoding p115RhoGEF-GFP. 48 h after transfection, cells were fixed and stained with a monoclonal anti-HA antibody, followed by Alexa-594-conjugated anti-mouse antibody, to stain for the HA epitope tagged α₁₃ constructs (anti-HA panels). p115RhoGEF’s localization was determined by visualization of the intrinsic fluorescence of GFP (GFP panels), and DAPI staining was used to identify nuclei (DAPI panels). Three color merged images show α₁₃ (red), p115RhoGEF-GFP (green) and nuclei (blue) (Merged panels). A representative image of more than 100 cells analyzed in three separate experiments is shown.