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. 2019 Dec 12;9:18919. doi: 10.1038/s41598-019-55364-8

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© The Author(s) 2019

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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Effects of SCH on axodendritic morphogenesis in hippocampal neurons. Neurons were grown on the same culture conditions as indicated in Fig. 1 for 3–5 days. Neurons were then fixed and double-immunostained for ankyrin G (red) and α-tubulin (green). Ankyrin G is characteristically concentrated at axon initial segment (AIS; indicated by arrows). Representative fluorescent images from cultures at DIV3 or 5 to visualize the cell morphology. Scale bar, 60 μm, applies to all images (a). Morphometric analysis of DIV3~5 neurons for dendritic development: the number of primary dendrites (b), the length of primary dendrites (c), the number of dendritic branches (d) and the length of dendritic branches (e). Sholl analysis for dendritic intersections (f) and branch points (g) in DIV5 neurons. Morphometric analysis of axonal development: length of axonal shaft (h), axonal collateral branch (i) and the total length of axonal collateral branch (j) in orders. Sholl image with reconstructed neurons, which does not represent any of the experimental data (k). Sholl analysis for axonal intersections (l), and axonal collateral branch points (m). Bars represent the mean ± SEM (n >15 neurons). Data points represent the mean ± SEM (n = 15 neurons). Statistical significance compared to vehicle: *p < 0.05, **p < 0.01 and ***p < 0.001 (Student’s t-test).