FIGURE 2:

Recruitment of Myo1E-GFP when clathrin is being internalized. (A) A low-magnification TIRFM view of Swiss 3T3 cells stably expressing DsRed-clathrin light chain a (CLTA) and transiently expressing Myo1E-GFP. Right, a kymograph of the peripheral region of the same cell, showing recruitment of Myo1E to CCPs at the time of internalization. Bar, 10 μm. (B) Representative temporal recruitment profile of Myo1E-GFP and DsRed-CLTA in Swiss 3T3 cells. Myo1E-GFP is recruited concomitant with clathrin internalization. (C) Montage and (D) kymograph show the recruitment of Myo1E-GFP at the end of the lifetime of DsRed-CLTA. (E) Particle tracking of Myo1E-GFP shows a characteristic vectoral movement at the CCP with reference to CLTA. Bar, 100 nm. (F) Representative temporal recruitment profiles for GFP-tagged dynamin, N-WASP, and actin-marker utrophin (calponin-homology domain) in Swiss 3T3 cells stably expressing DsRed-CLTA (left). Images were captured at 2-s intervals. Representative temporal recruitment profiles of tagged dynamin, WIP, and WIRE in Swiss 3T3 cells expressing Myo1E-GFP (right). Images were captured at 1-s intervals.