Figure 3.

S277A GRASP65 FL inhibits Golgi orientation. (A) GRASP65 FL or S277A GRASP65 FL cDNA were microinjected into serum-starved cells at the wound edge. Proteins were expressed for 2 h before inducing polarization with LPA for 90 min. Cells were then labeled with antibodies against GRASP65 (green) and GM130 (red), and DNA was stained with Hoechst (blue). Injected cells were identified by overexpressed GRASP65. Golgi were counted as oriented and marked as in Fig. 1 A. Bar, 10 μm. (B) Quantitation of A. Shown are mean ± SEM of n = 4 independent experiments with an average of 27 cells per condition, per experiment. *, P < 0.016. Red lines mark basal levels expected for random orientation of 33%.