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. 2020 May 22;11:2565. doi: 10.1038/s41467-020-16328-z

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Measurement of the rotational pitch of speckled microtubules gliding on a reflective silicon substrate coated with GFP-Ncd via anti-GFP antibodies bound to Fab fragments. Due to fluorescence interference contrast (FLIC), the recorded intensities of the asymmetric speckles change as a function of height above the substrate. Rotational information of gliding speckled microtubules is interpreted from the periodic variation in the recorded intensity of the speckles on the microtubule (vertical scale bar: 10 μm; horizontal scale bar: 40 s in the example kymograph, FLIC intensity profile over time for one of the speckles indicated by the green line in the kymograph). The rotational pitch of this gliding microtubule is about 1 μm. b The rotational pitch of the gliding microtubules was dependent on the surface density of motors. The motor density was set by changing the Fab fragment dilution (Fab fragment stock concentration of 2 mg/mL; diluted 50×, 20×, 5×, and 2×; N = 25, 45, 69, and 24 events, respectively) and ranged between 0.5 and 2.0 μm. c FLIC-based measurement of the rotational pitch of speckled transport microtubules sliding on surface-immobilized template microtubules (vertical scale bar: 10 μm; horizontal scale bar: 40 s in the example kymograph, FLIC intensity profile over time for one of the speckles indicated by the purple line in the kymograph). The rotational pitch of this sliding transport microtubule is about 0.8 μm (see also Supplementary Movie 3). d The rotational pitch of the sliding transport microtubules was dependent on the motor concentration (0.6, 2.4, and 6 nM; N = 7, 16, and 29 events, respectively) and ranged between 0.5 and 2 μm. All the rotational pitch distributions in (b) and (d) are significantly different. Supplementary Fig. 4C provides the corresponding p values obtained from two-sample two-sided Mann–Whitney U-tests. For the boxplot description, see Methods.