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. 2018 Jan 24;8:1552. doi: 10.1038/s41598-018-19823-y

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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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aSyn distribution between soluble (TBS), membrane bound (TrX) and insoluble (SDS) fractions in the presence of PREP overexpression. HEK-293 and PREPko cells were transfected with aSyn or aSyn + PREP and treated with oxidative stress for 48 hrs. Analyses of aSyn + GFP was not performed as the combination was very toxic to the cells but has been included in the WB images. (A–C), p129-aSyn levels significantly correlated with plasmid transfection and stress in PREPko and HEK-293 cells. (A) In TBS fraction, p129-aSyn was reduced in PREPko cells after aSyn + PREP transection. (B) Interestingly, combination of aSyn + PREP reduced considerably the levels of TrX p129-aSyn. (C) Moreover, oxidative stress significantly reduced the levels of SDS soluble p129-aSyn in both cell lines. (D–F), similar changes were seen in total aSyn levels, (E) although the decrease in TrX aSyn after aSyn + PREP transfection was not as notable as in p129-aSyn. (F) Moreover, in SDS soluble aSyn, there was a significant reduction after oxidative stress in both cell lines, and also aSyn + PREP transfection lowered the SDS-aSyn expression. Full-length blots are presented in Supplementary Figure S1. Bars represent mean ± SEM, 2-way ANOVA.