Rho GTPases regulate cell movement by controlling polarity, protrusion, and adhesion. In a wound healing assay, primary embryo fibroblasts were studied to understand the roles of Rho, Rac, and Cdc42 in cell movement. Rac is essential for lamellipodia protrusion and forward movement, while Cdc42 maintains cell polarity, including the localization of lamellipodial activity to the leading edge and reorientation of the Golgi apparatus. Rho is required for cell adhesion, and Ras regulates focal adhesion and stress fiber turnover, which is essential for cell movement. These four GTPases work together to promote cell movement. The study shows that Rac is essential for leading edge protrusions, Cdc42 is required for cell polarity, and Rho is necessary for cell adhesion. Ras is also essential for focal adhesion turnover during migration. The findings highlight the importance of Rho GTPases in regulating cell movement during development and tissue repair.Rho GTPases regulate cell movement by controlling polarity, protrusion, and adhesion. In a wound healing assay, primary embryo fibroblasts were studied to understand the roles of Rho, Rac, and Cdc42 in cell movement. Rac is essential for lamellipodia protrusion and forward movement, while Cdc42 maintains cell polarity, including the localization of lamellipodial activity to the leading edge and reorientation of the Golgi apparatus. Rho is required for cell adhesion, and Ras regulates focal adhesion and stress fiber turnover, which is essential for cell movement. These four GTPases work together to promote cell movement. The study shows that Rac is essential for leading edge protrusions, Cdc42 is required for cell polarity, and Rho is necessary for cell adhesion. Ras is also essential for focal adhesion turnover during migration. The findings highlight the importance of Rho GTPases in regulating cell movement during development and tissue repair.