Fig. 3.

Distinct modes of membrane bending observed by pol-TIRF. a–c Kymographs of clathrin-Tq2, membrane bending (P/S), dynamin-eGFP, and corresponding intensity traces for clathrin (cyan) and P/S (red) for the three classes of membrane bending observed by pol-TIRF. The colored lines indicate the lifetime for each event, the dashed red line highlights the start of P/S event. a Class 1: clathrin and P/S signals proceed together indicating membrane bending as clathrin assembles. b Class 2: clathrin signal plateaus prior to the start of P/S indicating all required clathrin was present as a flat sheet. c Class 3: clathrin assembles prior to P/S signal, but new clathrin was recruited as the membrane bends and the vesicle is formed. d The clathrin lifetime histogram of each class from an average of four cells (N = 481 tracks total)