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. 2021 Aug 19;32(18):1641–1653. doi: 10.1091/mbc.E20-10-0685

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© 2021 Rey-Suarez et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 2: — Microtubule growth rates are differentially modulated across different regions and by Arp2/3- and formin-mediated actin architectures. (A) Representative two-color image of a Jurkat cell transiently transfected with tdTomato-F-tractin (red) and EGFP-EB3 (green). Scale bar is 5 μm. Higher magnification views at different time points of the region enclosed by the white square in the left-hand image are shown on the right. The white arrow points to a microtubule tip, whose movement is drastically reduced once it reaches the actin-rich distal region. Scale bar is 3 μm. (B) EB3 tracks from a different cell than the one shown in A, color coded for the region of the synapse where they occurred: central (black), peripheral (blue), and distal (red). (C) Comparison of instantaneous speeds of EB3 across the three regions considered. N = 11 cells. (D) Cumulative distribution of instantaneous angle differences of EB3 tracks as a measure of directionality persistence, defined as the angle difference between two consecutive displacements, in different regions. (E) Cumulative distribution of radial angles calculated from interframe displacements of EB3 tracks for the different regions. (F) Cumulative distribution of instantaneous EB3 speeds in the distal region classified as radial (radial angle ≤ 45°) or nonradial (radial angle > 45°). (G) Box plot of EB3 instantaneous speeds measured in the peripheral region cells treated with the actin nucleation inhibitor CK666 (Arp2/3) or SMIFH2 (formin) compared with DMSO control. N = 11 cells for DMSO, N = 11 for CK666 and N = 16 for SMIFH2. (H) Cumulative distribution of radial angles in the distal region for cells treated with CK666 or SMIFH2 compared with DMSO. Significance of differences was tested using the Kruskal–Wallis test (***p < 0.001, **p < 0.01, *p < 0.05). Number of data points: C, Cen 12930, Per 23962, Dist 9579. D and E, Cen 11563, Per 21964, Dist 8206. F, Rad 6047, Non Rad 2159. G, DMSO 23962, CK666 19909, SMIFH2 21128. H, DMSO 8206, CK666 17316, SMIFH2 9186.