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. 2018 Apr 24;9:1623. doi: 10.1038/s41467-018-03940-3

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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. 1 — Three-dimensional morphology of astrocytes in layers I–VI of the mouse somatosensory cortex as revealed by visualization of microtubule and whole-cell structures. a 3D reconstruction of astrocyte morphology in the Glast-EMTB-GFP mouse brain (P60) rendered transparent by the CUBIC technique. 3D structures and traces are shown in the left panels, and individual traces in the right. CC, corpus callosum. Scale bars, 250 μm (left panels), 50 μm (right panels). b Representative confocal image of sparsely labeled astrocytes in Glast-EMTB-GFP;Glast-CreER^T2;Rosa-CAG-LSL-tdTomato mice (P65) injected with a low dose of tamoxifen at P60 (left), and representative 3D projection images of astrocytes in each layer (right). Arrowhead indicates GFP⁺tdTomato⁺ cell. Scale bars, 250 μm (left panel), 50 μm (right panels). c, d Quantification of territorial volume (c) and the angle of orientation relative to the brain surface (d) for astrocytes in each layer. The data are shown for 116 cells from five brains, with the red bars indicating median values. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way ANOVA followed by Bonferroni’s test). e 3D Sholl analysis for microtubule structure of sparsely labeled individual astrocytes in layers II/III and VI of Glast-EMTB-GFP;Glast-CreER^T2;Rosa-CAG-LSL-tdTomato mice injected with a low dose of tamoxifen at P60. The data are means (layer II/III astrocytes, n = 27 cells from five brains; layer VI astrocytes, n = 16 cells from five brains). *P < 0.05, **P < 0.01, ***P < 0.001 versus corresponding values for layer VI astrocytes (two-way ANOVA followed by Bonferroni’s test)