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. 2018 Feb 9;8:2717. doi: 10.1038/s41598-018-21252-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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Activation of orexin neurons by CHA. (A) Square field denotes counting area for immunoreactivity. (B) The number of c-Fos positive cells among 4 groups of the mice: WT mice treated with vehicle, WT mice treated with 0.02 nmol of CHA, ORX-AB mice treated with low dose (0.02 nmol) of CHA, and ORX-AB mice treated with high dose (0.2 nmol) of CHA. 2-way ANOVA indicated there was statistical difference between doses (F_1,20 = 7.89, p < 0.0108) and between genotypes (F_1,20 = 19.86, p = 0.0002). P values were calculated using the Holm-Sidak multiple comparison test. (C) Percentage of the number of double positive cells among orexin immuno-positive cells. P value was calculated using the Mann-Whitney U-test. In (B) and (C), horizontal lines indicate mean and SEM of 6 animals. Keys for the brain structures: DMH, dorsomedial hypothalamus; f, fornix; LHA, lateral hypothalamic area; mt, mammillothalamic tract; PeF, perifornical area.