Fig. 3. Kinetics of uncoating of clathrin-coated pits by Hsc70-auxilin-ATP mixture. (A) Uncoating mixture composed of Hsc70 (2 µM), auxilin (0.2 µM) and ATP (1 mM) was added to the permeabilized cells and imaged at 30-second intervals. (B) The decrease in fluorescence intensity of three different regions of interest (ROI) (circles, numbered 1-3) for the time course in A is plotted against time. Data were normalized to the maximal initial fluorescence in each plot. Bar, 10 µm.