Figure 6.2.

Dominant-negative Pak1 accumulates in focal contacts in scrape-wounded epithelial cells. Image represents MDCK cells, which express wild type or a dominant-negative (Pak1-K299R, kinase-dead) under control of a tetracycline-regulatable promoter (Zegers et al., 2003a).Monolayers of MDCK cells expressing these HA-tagged forms of Pak1 were scrape wounded. The next day, cells were fixed and stained using antibodies against the HA-tag and the focal contact marker vinculin. Note that wild type Pak1 localized to focal contacts to a limited extent, and localization is restricted to very peripheral focal contacts. In contrast, kinase-dead Pak1 accumulates at focal contacts and is found both peripheral and more distally within lamellipodia.