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Fig. 3. Actin assembly at CCS is Arp2/3 complex dependent. TIRFM images of N-WASP^flox/flox fibroblasts transiently co-expressing mRFP-clathrin light chain A (red in merges), EGFP-actin (green in merged images), and either WA-Scar1 or W-Scar1. Note the disappearance of CCS in cells expressing WA without any detectable coincident actin polymerization (asterisks) and the apparent actin assembly at disappearing CCS in the W-expressing control (arrowheads). Time is given in seconds; scale bar: 1 µm. The quantifications (bottom) of actin recruitments during CCS internalization revealed a complete abolition upon WA over-expression (0%, n=16, 110 events) and levels comparable to wild-type cells (Fig. 1D) in W-expressers (69.9±2.7%, n=16, 223 events).

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