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. 2020 Jun 1;11(6):411. doi: 10.1038/s41419-020-2615-9

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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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Fig. 2 — a Immunoblotting analysis of GFAP, BDNF, and tubulin as a loading control in WT and pGFAP-BDNF mice treated with vehicle or pilocarpine. b Densitometry quantification of GFAP results as in a. Statistical analysis revealed a significant increase on GFAP protein levels in groups of mice treated with pilocarpine (two-way ANOVA, variable treatment: F_(1,22) = 17.44, p < 0.001). c Densitometry quantification of the mature form of BDNF (mBDNF) results as in a. Statistical analysis revealed a significant increase on mBDNF protein levels (two-way ANOVA variable treatment: F_(1,26) = 16.89, p < 0.001) specifically in pGFAP-BDNF mice treated with pilocarpine (post hoc test: p < 0.001). d Densitometry quantification of pro-BDNF results as in a. Statistical analysis revealed a significant increase on pro-BDNF protein levels (two-way ANOVA variable treatment: F_(1,20) = 17.50, p < 0.001) in both WT and pGFAP-BDNF mice treated with pilocarpine. e Representative image showing GFAP immunofluorescence in the hippocampus. Scale bar, 500 μm. f Representative immunofluorescence images showing GFAP and BDNF immunoreactivity in the hippocampus of WT and pGFAP-BDNF mice treated with vehicle or pilocarpine. Scale bar, 20 µm. g Quantification of astrocytic BDNF immunoreactivity. Statistical analysis revealed a significant increase in the astrocytic BNDF (two-way ANOVA interaction: F_(1,149) = 16.37, p < 0.0001; variable treatment: F_(1,149) = 235.8, p < 0.0001; variable genotype: F_(1,149) = 20.55, p < 0.0001). Data were analyzed by two-way ANOVA followed by Bonferroni’s test. *p < 0.05, **p < 0.01, ***p < 0.001 compared to Veh. ^$$$p < 0.001 compared to WT-Pilo. All mice were used at 10 weeks of age and were males.