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. 2017 Sep 14;7:11547. doi: 10.1038/s41598-017-11739-3

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© The Author(s) 2017

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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Interaction of HDAC6 with microtubules visualized by TIRF microscopy. (a) HDAC6 interacts directly with microtubules. Cy5-labeled microtubules were immobilized on a glass coverslip surface. Micrographs show microtubules (red) in the presence of 500 nM HDAC8-FITC (green; negative control, upper panel) or 475 nM FITC-labeled HDAC6 (green; lower panel). In the presence of FITC-labeled HDAC6, fluorescent signals of microtubules and HDAC6 co-localize along the whole length of microtubules (yellow). (b,c) HDAC6-GFP binds uniformly along the whole length of the microtubules. GFP-HDAC6 fusion was added to surface-immobilized microtubules and visualized in the 488 channel using TIRF microscopy. Uniform increase in GFP fluorescence intensity was observed along the whole length of microtubules. The kymograph in Panel B depicts averaged (background subtracted) GFP fluorescence intensity of 4 microtubules of the same length (approximately 12 µm) after the addition of HDAC6-GFP in the timespan from 0 to 60 seconds. The right panel shows the averaged GFP signal (±s.d.) over time at the edge of the microtubules (orange line) and in the middle (blue line). The regions (width of 1.5 µm) used for the averaging are indicated in the kymograph (b) by orange and blue rectangles.