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. 2020 Sep 4;8:155. doi: 10.1186/s40478-020-01030-4

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — C9orf72 regulates the expression of a longevity gene, Klotho, and is required for adult neurogenesis in the hippocampus. a Schematic for transcriptomic analysis of the hippocampus from wild type and C9orf72 knockout mice. b Total of 60 differentially expressed genes (DEGs) were identified. Among them, 48 and 12 genes are up- and down-regulated, respectively. Furthermore, 14 of 48 (29.1%) up- and 7 of 12 (58.3%) down-regulated genes belong to the noncoding RNAs (magenta). c Gene otology analysis of up-regulated DEGs. d Age-dependent deregulation of Klotho expression in C9orf72 knockout mice. Schematic of two Klotho isoforms due to the alternative usage of exon 3. RNAs were extracted from wild type and C9orf72 knockout mice, reverse transcribed and quantified using primers specific for isoform 1 and 2 of Klotho gene. Sub-panel i, ii, and iii are qRT-PCR results for 3, 6, and 12-month animals. *p < 0.05, ***p < 0.0001. (di) 3 months, KL-L: p = 0.0328; KL-S, p = 0.0476, (dii) 6 months, KL-L: p = 0.4734; KL-S, p = 0.9766, and (diii) 12 months, KL-L, p = 0.0002; KL-S, p = 0.0008. n = 3, per genotype, per timepoint. e Confocal images of Klotho protein in CA1 and DG region of wild type and C9orf72 knockout mice. Klotho immunoreactivity is increased at the dendritic region of CA1 and reduced in the granule cell layer of DG in the C9orf72 knockout mice. Scale bar is 20 μm. n = 3 per genotype. f Schematic of EdU-pulse chase experiment (left panel). g Confocal image of EdU/doublecortin (DCX) staining. Scale bar is 50 and 10 μm, respectively. h Quantification of EdU-positive cells. i Quantification of EdU/DCX-double positive cells in the DG region. (3–5 slices per animals, n = 3 per genotype, p < 0.05)