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. 2018 Jan 4;9:70. doi: 10.1038/s41467-017-01859-9

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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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Fig. 3 — MtDNA loss in astrocytes causes massive astrocyte activation. a, b Astrocytes (GFAP, red) in somatosensory cortex (a) and in hippocampus, CA1 area (b) of 7–8-month-old mice and 3–4 months as shown in inset of (a). In b, boxed area is enlarged on right. c Astrocytes (GFAP, red) immuno-co-stained with reactive astrocyte marker (Nestin, green). Insets enlarge the boxed area. Arrows show the same cells. d Astrocytic marker Aldh1l1 immunohistochemistry, counterstaining hematoxylin. TwKO^astro show intensive vacuolization. Brain graphs show sampling site. Fluorescence figures are counterstained with DAPI (blue). Bar graphs on right hand side: corresponding quantifications of GFAP⁺ cells (cortex and hippocampus); GFAP⁺Nestin⁺/GFAP⁺, and Aldh1l1-positive cells in somatosensory cortex. The data are presented as mean and error bars indicate standard deviation. Statistical significance: *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars: 20 μm (a), 50 μm (b–d)