Fig. 3. Targeted PAI-1 downregulation inhibits wound-stimulated cell motility. RK cells were transfected with a non-insert-bearing vector (Rc/CMV) or with constructs in which full-length PAI-1 coding sequences were cloned in antisense (Rc/CMVIAP) or sense (Rc/CMVPAI) orientation (A). Cultures were grown to confluency and scrape-wounded. Extent of repair-associated migration (% wound closure) was measured over a 24 hour period (B). There was no difference in stimulated motility among Rc/CMV- or Rc/CMVPAI-transfectants compared with non-transfected controls (RK). The rate of monolayer scrape repair by Rc/CMVIAP (antisense PAI-1)-transfected cells, in contrast, was significantly impaired (asterisk) relative to control RK cultures or to sense (Rc/CMVPAI) or empty vector (Rc/CMV) transfectants. Data plotted is mean±standard deviation of three independent wound repair determinations. Inset in B is a western blot of PAI-1 levels in the various cell types at the 24 hour time point illustrating downregulation of PAI-1 expression in the Rc/CMVIAP transfectants.