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. 2024 Apr 9;14:8346. doi: 10.1038/s41598-024-59061-z

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© The Author(s) 2024

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 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/.

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Diurnal variation of nest-building behavior and nest-building assay. (a–d) Representative pictures of nests at ZT6 (a), ZT12 (b), ZT16 (c), and ZT19 (d). (e) The proportion of nest height scales that was evaluated at that point in time every hour (ZT10-12, 8 mice; ZT13-1, 6 mice; ZT2-3, 8 mice; ZT4-9, 5 mice). (f) Relative time spent in nest-building (ZT10-12, 8 mice; ZT12-20, 6 mice; ZT20-22, 5 mice; ZT22-2, 6 mice; ZT2-3, 8 mice; ZT3-5, 4 mice and ZT5-10, 5 mice). (g) Diagram of the nest-building assay. After single-housed for five days or longer, the nest was removed and fresh nest material was immediately introduced to observe nest-building behavior. (h) Accumulated time manipulating nest material from the point of introducing the fresh nest material (ZT6-male, n = 5; ZT6-female, n = 5; ZT12-male, n = 5). Data are shown as the mean ± s.e.m.