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. 2018 Oct 1;217(10):3531–3544. doi: 10.1083/jcb.201802039

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© 2018 Wu and Bezanilla

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Figure 3. — Microtubules confine class II formins and the actin cluster to the cell apex. (A) A WT cell expressing Lifeact-mEGFP (green) and For2A-mCherry (magenta). Images are single focal planes acquired every 0.27 s. Bar, 5 µm. (B) Kymograph generated from the time lapse acquisition shown in A along the yellow dashed line. Bars, 5 µm (vertical) and 20 s (horizontal). (C) A 5-µm diameter circle near the cell tip was tracked using TrackMate, and the mean intensity of Lifeact-mEGFP and For2A-mCherry signals over time are shown in the plot. (D) The same cell in A after 12.5 µM oryzalin was added to the microfluidic device. Images are single focal planes acquired every second. Bar, 5 µm. (E) Foci enriched with For2A-mCherry were tracked using TrackMate and the mean intensity of Lifeact-mEGFP and For2A-mCherry signals were plotted over time. Numbers above the peaks in the plot correspond to the bursts of actin polymerization indicated in D. Time stamps represent min:s. A.U., arbitrary units. See also Video 5.