Direct imaging of clathrin and β1 integrin during focal adhesion disassembly. (a) TIRF imaging of GFP–β1 integrin during MT-induced focal adhesion disassembly. Time is in minutes:seconds after nocodazole washout. The boxed region is shown magnified at various time points in the panels on the right. (b and c) Montages from TIRF imaging of DsRed-clathrin and GFP–β1 integrin during focal adhesion disassembly. Time is in minutes:seconds. The boxed areas are depicted in the panels on the right. In the merged panels, arrowheads point to clathrin/β1 integrin puncta that colocalize and then simultaneously disappear. (d) Quantification of the behavior of β1 integrin–associated clathrin puncta. The histogram represents data from 30 focal adhesions in four different videos, in which 50 individual clathrin puncta were analyzed during focal adhesion (FA) disassembly. Error bars are SEM. Bars: (a) 10 µm; (b and c) 2 µm.