Effects of Na/K-ATPase knockdown on the mobility of Cav1-YFP–positive vesicles. Control (P-11) and knockdown (TCN23-19) cells were transfected with Cav1-YFP, and Cav1-YFP–positive vesicles moving underneath or away from the plasma membrane were monitored using time-lapse confocal microscopy. (A) Trajectories of the moving Cav1-YFP–positive vesicles in control, knockdown cells, as well as in the knockdown cells that were treated with 5 μM of nocodazole (Noc) or 2 μM of PP2. The moving path of each moving vesicle was shown as a line and ended with a dot. The images are the overlay of the trajectory with the synergized snapshots from Videos 1, 3, 5, and 7. (B) The mean square distance (MSD) each Cav1-YFP–positive vesicle traveled between each frame was measured, and the mean ± SE from at least 30 measurements is shown in the graph. (C) The average of maximal range of each vesicle traveled was calculated and shown in the bar graph. Values are mean ± SE of 16 separate experiments of each cell line. **, P < 0.01 between groups as indicated. Bar, 10 μm.