Laser ablation has emerged as a useful technique to study the mechanical properties of the cytoskeleton in living cells. Laser ablation perturbs the force balance in the cytoskeleton, resulting in a dynamic response which can be imaged. Quantitative measurement of the dynamic response allows the testing of mechanical theories of the cytoskeleton in living cells. This review discusses recent work in applying laser ablation to study stress fiber and microtubule mechanics in living endothelial cells. These studies reveal that molecular motors are major determinants of the mechanical properties of the cytoskeleton in cells.