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. 2019 Sep 18;9:235. doi: 10.1038/s41398-019-0569-4

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© The Author(s) 2019

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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Fig. 3 — Two-way ANOVA with Tukey post hoc test was performed (males (M): N.Ch. Adnp^+/+ N = 12–26; N.Ch. Adnp^+/− N = 12–18; +Ch. Adnp^+/+ N = 21; +Ch. Adnp^+/− N = 15; +Ch. Adnp^+/+ PACAP N = 14; +Ch. Adnp^+/− PACAP N = 12; females (F): N.Ch. Adnp^+/+ N = 18; N.Ch. Adnp^+/− N = 12; +Ch. Adnp^+/+ N = 19; +Ch. Adnp^+/− N = 17; +Ch. Adnp^+/+ PACAP N = 24; +Ch. Adnp^+/− PACAP N = 17). For gene expression analysis, results are presented as 2^−ΔCT, normalized to Hprt. For correlation testing, if both plotted data sets were normally distributed, a Pearson’s correlation analysis was performed. If at least one of the data sets was not normally distributed, a Spearman’s correlation was performed. a Splenic Adnp expression in vehicle-treated males, significant main genotype effect (F(1,76) = 5.664, p = 0.020) was found, with significant differences between non-challenged Adnp^+/+ and Adnp^+/− mice (*p < 0.05). b Splenic Adnp expression in stressed-challenged females: main genotype (F(1,73) = 15.336, p < 0.001), treatment (F(1,73) = 19.029, p < 0.001), and interaction (F(1,73) = 23.179, p < 0.001) effects were found, with significant differences between challenged Adnp^+/+ and Adnp^+/− mice (***p < 0.001), as well as challenged PACAP- vs. vehicle-treated Adnp^+/+ mice (***p < 0.001). In vehicle-treated females, significant main genotype (F(1,62) = 26.464, p < 0.001) and interaction (F(1,62) = 38.150, p < 0.001) effects was found, with significant differences between non-challenged Adnp^+/+ and Adnp^+/− mice (***p < 0.001), challenged vs. non-challenged Adnp^+/− mice (***p < 0.001), and challenged vs. non-challenged Adnp^+/+ mice (***p < 0.001). c In non-challenged males (N = 12), splenic Adnp gene expression was positively correlated with the D2 score in the novel object recognition test (Pearson’s correlation, r = 0.637, *p < 0.05)