Fig. 8. PDZ-RhoGEF is required for LPA induction of adhesion movement. (A) Analysis of adhesion movement in response to LPA. Control Rat2 cells and Rat2 cells treated with PDZ-RhoGEF siRNA that stably expressed paxillin-GFP were plated on fibronectin-coated dishes in the presence of medium containing 10% serum. The following day, cells were serum starved for 12-16 hours and then stimulated with 2 µM LPA. Adhesion dynamic was analyzed using TIRF microscopy as described in Materials and Methods. Cells were filmed for 21 minutes before the addition of 2 µM LPA and 27 minutes after the addition of 2 µM LPA. (B) Quantification of dynamic adhesions (arrows indicate representative adhesions). The experiment was performed twice with five to seven cells analyzed per condition.