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. 2018 May 9;8:7369. doi: 10.1038/s41598-018-25331-w

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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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Immunofluorescence detection of BrdU-, NeuN-, and Zif268-positive cells. (A) Representative epifluorescence images from four sections used for the analyses. The sections were immunostained for NeuN. One in every 12 of the 40-µm-thick sections was selected, and the selected sections were designated #1–4. Scale bar: 500 µm. (B) An example of confocal image tiles, consisting of 12 images (512 × 512 pixels each) covering the whole granule cell layer. Blue: NeuN, red: Zif268, green: BrdU. Scale bar: 200 µm. (C) A confocal image showing BrdU/NeuN/Zif268-triple-positive cells (arrowheads). Scale bar: 30 µm.