Figure 6. Effects of Rho kinase inhibitor on LPA- and thrombin-induced cytoskeletal reorganization

Pretreatment with Rho kinase inhibitor (Y-27632) prevents LPA and thrombin induced formation of actin stress fibers and focal adhesions in human lens epithelial cells. As compared to cells stimulated with LPA (A) and thrombin (C) alone, cells pretreated with Y-27632 (5 μM for 20 min) fail to respond to these growth factors (B and D), as evidenced by the lack of actin stress fiber and focal adhesion formation. These observations indicate involvement of the Rho/Rho kinase pathway in mediating the effects of LPA and thrombin on cytoskeletal reorganization in lens cells. Cells were double-labeled for actin stress fibers (FITC-phalloidin) and focal adhesions (vinculin primary antibody with TRITC-conjugated secondary antibody). Punctate yellow-orange staining at the leading edges of cell membrane represents staining of focal adhesions (shown with arrows).