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. 2015 Dec 15;26(25):4646–4659. doi: 10.1091/mbc.E15-09-0626

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© 2015 Young, Heimsath, and Higgs. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 2: — Arp2/3 complex inhibition reduces formin-mediated filopodia in adherent cells. (A) Representative micrographs of U2OS cells transfected with GFP-mDia2-FFC and treated for 2 h with 200 μM CK689 (left) or 200 μM CK666 (right). Actin is stained with TRITC-phalloidin (red), and GFP-mDia2-FFC is green. Arrowheads indicate mDia2 enriched at filopodial tips. Filopodia number in insets: GFP/DMSO, 0; mDia2/CK689, 10; and mDia2/CK666, 5. Examples of untreated U2OS cells transfected with GFP alone in Supplemental Figure S3. (B) Quantification of percentage of transfected cells displaying filopodia after the indicated treatment. (C) Quantification of filopodia number on transfected cells treated as indicated. Results are pooled from three or four independent experiments, 24–37 cells (FMNL3) and 92–141 cells (mDia2). (D) Representative micrographs of NIH 3T3 cells transfected with GFP-FMNL3-FF and then treated for 2 h with DMSO (left) or 200 μM CK666 (right). Actin is stained with TRITC-phalloidin (red) and DNA with DAPI (blue), and GFP-FMNL3-FF is green. Arrowheads indicate FMNL3 enriched at filopodial tips. Filopodia numbers in insets: DMSO, 9; and CK666, 2. Scale bar, 10 μm (main image) and 5 μm (insets). (E) Quantification of percentage of transfected cells displaying filopodia after indicated treatment. (F) Quantification of filopodia number on each transfected cell after the indicated treatment. Results are pooled from three to five independent experiments; 48–118 cells. Scale bars: 10 μm (main image) and 5 μm (insets). Error bars, SD. ***p < 0.001.