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. 2020 Jul 3;10:11017. doi: 10.1038/s41598-020-67291-0

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© The Author(s) 2020

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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NBCn1 is downregulated in the brains of mice after chronic alcohol consumption. (A) Comparison of NBCn1 expression in the brains of pair-fed vs. alcohol-fed mice. C57BL/6 mice were fed with 20% alcohol or water in the two bottle free choice procedure for 6 weeks and NBCn1 immunostaining was compared (n = 4 mice/group). The scale bar in the upper image is 1 mm and applies to the bottom image. (B) Comparison of NBCn1 immunostaining in the pyramidal CA3 region (Hc), caudate putamen (CP), and nucleus accumbens (NAc). The regions were identified using the Allen Brain Atlas (https://mouse.brain-map.org/static/atlas). The scale bar in the upper left image is 20 μm and applies to all images. (C) Quantitation of NBCn1 immunostaining. NBCn1-immunostained neurons were randomly selected and quantitated using ImageJ software (n = 10/region). Cb, cerebellum; Cx, cortex; Hc, hippocampus; CP, caudate putamen; NAc, nucleus accumbens; SC, superior colliculus.