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. 2017 May 12;7:1876. doi: 10.1038/s41598-017-01963-2

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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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Non-phosphorylated ZBP1 granules are more motile. (a) Representative kymographs showing motility of EGFP-ZBP1 and EGFP-ZBP1_S181A granules in dendrites. (b,c,d) Relative numbers of ZBP1 and ZBP1_S181A granule movements. Granule movements were quantified, normalized per 10 μm of the dendrite length, and then normalized to the mean number of anterograde movements in the wildtype ZBP1 variant for each experiment, respectively. The data in the columns are expressed as a ratio to the mean number of movements. (e) Granule velocity was normalized to the mean anterograde velocity in the wildtype ZBP1 variant for each experiment, respectively. ***p < 0.001, *p < 0.1 (Student’s t-test). The data were analyzed from five neurons per group per experiment, with three independent experiments. Total raw number of directed anterograde or retrograde movements per group per experiment ranged from 15 to 57, and the number of oscillatory anterograde or retrograde movements per group per experiment ranged from 99 to 245 (for complete data see Table S1). The data are expressed as mean ± SD. (f) Western blot analysis of KIF5A-GFP bound to HA-BIO-ZBP1 or HA-BIO-ZBP1_S181A immunoprecipitated from Neuro2a cells transfected as indicated. Cropped blots are presented (for full length blots see Supplementary Information).