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. 2017 Apr 28;7:1277. doi: 10.1038/s41598-017-01215-3

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© The Author(s) 2017

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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Changes in human astrocytes intracellularly accumulated and extracellularly shed amounts of sAPPα according to treatments. (a) Cumulative curves of sAPPα shed into the medium by astrocytes as affected by treatments. Points on the curves are means ± SEM of 5 independent experiments. One-way ANOVA analysis of each complete data set: fAβ_25–35: F = 80.877, P < 0.001; fAβ_25–35 + NPS 2143: F = 35.365, P < 0.001; fAβ_25–35 + CMT: F = 50.882, P < 0.001; fAβ_25–35 + CMT + NPS 2143: F = 34.261, P < 0.001. Bonferroni t-test: comparisons of each treated group versus 0-h group (controls): *P < 0.05; pair-wise comparisons between corresponding treatments ± NPS 2143 at each time point: ^#P < 0.05. (b) Top. Typical immunoblot revealing the changes in intracellularly accumulated sAPPα according to treatments versus controls (0-h). LC, loading control. Blots have been cropped to size for clarity. Bottom. Densitometric evaluations of intracellular sAPPα bands. Points on the curves are means ± SEM of 3 independent experiments, with 0-h values normalized as 1.0. One-way ANOVA analysis of each complete data set: fAβ_25–35 ± NPS 2143: F = 280.483, P < 0.001; fAβ_25–35 + CMT ± NPS2143: F = 134.239, P < 0.001. Bonferroni t-test: comparisons of each treated group versus 0-h group (controls): *P < 0.05; pair-wise comparisons between corresponding treatments ± NPS 2143 at each time point: ^#P < 0.05. (c) The alterations in total 0-to-72-h amounts of extracellularly shed and intracellular sAPPα according to treatments or no treatment (controls). Bars are means ± SEM of 3–5 independent experiments. One-way ANOVA analysis of the (i) shed sAPPα complete data set: F = 44.056, P < 0.001; (ii) intracellular sAPPα complete data set: F = 587.435, P < 0.001. Bonferroni t-test: comparisons of each treated group versus untreated group (controls): *P < 0.05; pair-wise comparisons between corresponding treatments ± NPS 2143 of both intracellular and shed values, respectively: ^#P < 0.05. (d) Changes in secreted Aβ₄₂/sAPPα ratio values according to treatments or no treatment (controls). Bars are means ± SEM of 5 separate experiments. One-way ANOVA analysis of the complete data set F = 22.123, P < 0.001. Bonferroni t-test: comparisons of each treated group versus 0-h (controls): *P < 0.05; pair-wise comparisons between corresponding treated group versus added NPS 2143 group: ^#P < 0.05.