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. 2018 Mar 14;9:1067. doi: 10.1038/s41467-018-03480-w

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© The Author(s) 2018

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. 5 — The GFP-OsKCH1/KCH2 chimera is a processive minus-end-directed motor. a Schematic diagrams of GFP-OsKCH1(292–744) and the GFP-OsKCH1/KCH2 chimera. GFP-OsKCH1/KCH2 is a derivative of GFP-OsKCH1(292–744) by replacing its endogenous CC2 with that from OsKCH2. b Photobleaching analyses of GFP-OsKCH1/KCH2. (Top) Example fluorescence intensity traces over time of individual GFP-OsKCH1/KCH2 molecules immobilized on the microtubules. (Bottom) Histogram of the photobleaching steps of GFP-OsKCH1/KCH2 (n = 214). c Example kymograph of individual GFP-OsKCH1/KCH2 molecules (green) moving processively toward the minus end of single polarity-marked microtubules (red). d Velocity histogram of single GFP-OsKCH1/KCH2 molecules. Red line indicates a Gaussian fit to the velocity histogram. e Run-length histogram of single GFP-OsKCH1/KCH2 molecules. Red line indicates a single-exponential fit to the run-length histogram. f Coomassie-stained SDS-PAGE of the microtubule co-sedimentation assay for (top) GFP-OsKCH1(292–744) and (bottom) GFP-OsKCH1/KCH2 in BRB80/25 mM KCl. Scale bars: 30 s (vertical) and 5 µm (horizontal)