Fig. 2. The time course of guanosine 5′-[γ-thio]triphosphate (GTPγS)–stimulated membrane translocation of RhoA and Rock-2 in rat aortic smooth muscle. Rat aortas were homogenized in lysis buffer before (t = 0 min) or 1, 5, 20, and 60 min after treatment with GTPγS (10⁻⁴m). Membrane translocation of RhoA and Rock-2 began immediately at 1 min, continued to increase at 5 min, and increased to the maximal level at 20 min, which was sustained up to 60 min after application of GTPγS. The translocation of Rho seemed to be more complete than that of Rock-2. The image was representative of four independent experiments. C = cytosolic fraction; M = membrane fraction.