Figure 3.

The focal adhesion localization of TarP requires its LDVBD domain and the host protein vinculin, but not FAK. A, WT or vinculin-knockout MEFs were transfected with different mTurquoise2-tagged TarP constructs (green) and imaged by confocal microscopy to evaluate colocalization with paxillin (red) at focal adhesions. Phalloidin was used to stain F-actin (blue). Cells were transfected for 20 h, at which time the cells were fixed and processed for immunofluorescence staining for paxillin. B, in a parallel experiment, WT and FAK-deficient MEFs were transfected to express FLAG-HA-LDVBD and stained for FLAG (green). Colocalization with paxillin (red) was assessed by confocal microscopy. C, ectopically expressed LDVBD localizes to β1-integrin and paxillin-positive focal adhesions. Scale bar, 10 μm.