Fig. 9. LPA induced adhesion movement requires ROCKI and ROCKII. (A) Western blot analysis of ROCKI and ROCKII protein expression in control NIH3T3 cells and NIH3T3 cells treated with shRNA targeting FAK, ROCKI or ROCKII (top panel). Micrographs show phase-contrast images of control NIH3T3 cells and NIH3T3 cells treated with shRNA targeting ROCKI or ROCKII. (B) Control NIH3T3 cells and NIH3T3 cells treated with shRNA targeting ROCKI or ROCKII were serum starved for 12 hours. Following starvation, cells were stimulated with 2 µM LPA and adhesion movement was visualized by TIRF microscopy. (C) Quantification of adhesion movement (as described in Materials and Methods) in unstimulated serum-starved cells and in serum-starved cells stimulated with LPA (arrows indicate representative mobile adhesions). The experiment was performed twice with six to eight cells analyzed per experimental condition. (D) Quantification of cellular elongation in LPA-stimulated control cells and ROCKII-shRNA-treated cells. The experiment was performed twice.