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. 2017 Jul 20;7:6024. doi: 10.1038/s41598-017-06243-7

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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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Nucleocytoplasmic translocation of HDAC9 is required for TC axon branching via MEF2 regulation. (a) Average number of branch points per axon. Thalamic cells were electroporated with only mCherry, HDAC9 co-electroporated with mCherry, nucHDAC9 co-electroporated with mCherry, and nucHDAC9ΔMEF2 co-electroporated with mCherry. N = 12, 17, 18 and 14 axons from 4–6 co-cultures for each condition, respectively. (b) Branch density per axon. (c) Total branch length per axon. (d) Branch tip length. Statistical analysis by Kruskal-Wallis test with Dunn correction comparing the 4 columns. *p < 0.05. ***p < 0.001. ****p < 0.0001. e. Laminar distribution of branching points according to their distance from the pial surface. Arrows indicate presumed layer 4. Statistical analysis by two-way ANOVA for whole column effects with Tukey’s multiple comparisons test confirmed a significant difference in laminar specificity between nucHDAC9 and control (p < 0.0001), while no significant difference was found between nucHDAC9ΔMEF2 and control or wild-type HDAC9.