Fig. 4. Time course of uncoating of clathrin puncta by Hsc70-auxilin-ATP mixture. (A,B) Fluorescence image (A) showing the circled GFP-clathrin puncta (1-5) used to obtain the time course of uncoating (B). Bar, 5 µm. (C) Distribution in uncoating time obtained from 200 puncta (five cells) uncoated by Hsc70, auxilin and ATP using the same concentrations as in Fig. 2. Puncta measured were no larger than 0.3 µm. Cells were imaged using the 63x, 1.4 NA objective.