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. 2022 Oct 28;12:18196. doi: 10.1038/s41598-022-23091-2

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© The Author(s) 2022

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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Full photomicrograph analyses of microglial morphology were not sensitive to differences between control and treatments groups. Photomicrograph-averaged Iba1 percent coverage masked a difference between control and treatment groups that photomicrograph-specific percent coverage detected. 3 brain slices per mouse and 3 photomicrographs per brain slice were used. (a) Mean percent coverage of Iba1 staining per animal. Treatment n = 12 mice, Control n = 13 mice. (b) Percent coverage of Iba1 staining, where individual data points (gray dots) were not averaged. Full photomicrograph skeletal analysis detected that microglia from the treatment group had fewer endpoints than the controls but did not detect differences in branch number or branch length. (c) Mean microglial branch length; (d) mean number of endpoints per cell per photomicrograph; (e) mean number of microglial cells per photomicrograph; and (f) mean number of microglial branches per cell per image. Individual data points are presented as gray dots. (d–f) Treatment n = 108 images (12 mice, 9 images/mouse), Control n = 117 images (13 mice, 9 images/mouse). Results are presented as point estimates with 95% confidence intervals, which were estimated from mixed effects models with a negative-binomial error distribution.