Dynamic and polarized recruitment of ERC1a near protrusive sites during migration. Dynamic and polarized recruitment of ERC1a near protrusive sites during migration. (A–C) Frames from supplementary material Movie 3 (A,B) and Movie 4 (C). (A) ERC1a-positive vesicle-like structures appear near the protrusive edge. The dashed line in each panel indicates the protruding edge at t = 0. Arrow and arrowhead show the position of two vesicles moving centripetally from the edge. (B) Inverted images showing the edge of the protruding lamellipodium and the position of the GFP–ERC1a-positive compartment at t = 0 and t = 560 s. Merge: color-coded distribution of the GFP–ERC1a-positive compartment at t = 0 (green) and t = 560 s (blue). Scale bars: 5 µm. (C) Frames from an ERC1a-positive cluster showing the dynamic nature of the compartment: yellow arrowheads show an example of ERC1a-positive structure with dynamic changes in morphology. Scale bar: 3 µm. (D) ERC1a-positive (ERC) clusters partially colocalize with caveolin-1 at the cell periphery (arrows), whereas no co-staining is observed with clathrin-positive endocytic vesicles detected with antibodies for the heavy chain (HC) of clathrin. Scale bars: 20 µm. Veronica Astro et al. J Cell Sci 2014;127:3862-3876 © 2014. Published by The Company of Biologists Ltd