Video 6.
Single-object in vitro disassembly of clathrin/AP2 coats and sCCVs. The still image imaged with TIRF at the beginning of the time series corresponds to clathrin/AP2 coats and sCCVs containing PtdIns(4,5)P2 together with either PtdIns(3)P or PtdIns(4)P distinguished by labeling with DiI (red) or DiD (blue) lipid dyes. Before the time series, the channels corresponding to clathrin LCa–Alexa Fluor 488 (green) were shifted 5 pixels with respect to the lipids. The time series follows the uncoating reaction and corresponds to the clathrin fluorescence signal as a function of 25 nM ΔPTEN-Aux1 added together with 1 µM Hsc70 and 5 mM ATP. Data were acquired at 1-s intervals for 150 s using three-color TIRF microscopy.