Video 1.
WASP and SCAR complex distribution in wild-type cells. (left) Wild-type AX3 cells were transfected with GFP-WASP and clathrin light chain–mRFP. Cells were seeded onto a glass-bottom dish and incubated overnight in low fluorescent medium to reduce autofluorescence. Medium was aspirated, and cells were overlaid with 1% agarose to ensure consistent contact with the glass substratum. Time-lapse images were captured at two frames per second (f/s) on a microscope (Eclipse TE2000-U) that was fitted with a custom TIRF condenser. (right) Wild-type AX3 cells were transfected with the SCAR complex marker HSPC300-GFP. Cells were developed on Petri dishes under phosphate buffer until the onset of cell streaming. Cells were harvested, transferred to a glass-bottom dish, and overlaid with a thin layer of 0.4% agarose to ensure consistent contact with the glass substratum. Time-lapse images were collected at one frame per second on a microscope (Eclipse TE2000-U) that was fitted with a custom TIRF condenser. A differential interference contrast (DIC) image of the cell is overlaid in blue.