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. 2019 Feb 12;10(2):133. doi: 10.1038/s41419-019-1364-0

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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. 1 — a To investigate the effect of mutant A30P α-synuclein expression on autophagy, midbrain dopaminergic neurons expressing A30P α-synuclein were harvested and subjected to immunoblot analysis. P62 was used as an indicator for assessing autophagic activity. The intensity of each immunoblot band was quantified by densitometric scanning. b The insoluble fractions from midbrain dopaminergic neurons transfected with A30P α-synuclein were solubilized and separated by SDS-PAGE. c mRNA extracted from midbrain dopaminergic neurons transfected with A30P α-synuclein was used to assess amounts of p62 transcript relative to β-actin by RT-PCR. d A cycloheximide (10 μg/ml) chase experiment demonstrated that the decay kinetics of p62 in A30P α-synuclein-expressing cells was significantly prolonged compared with that in control group cells. The densitometric ratio is presented in the right panel. e midbrain dopaminergic neurons expressing A30P α-synuclein were treated or not with 3-MA (10 mM) for 24 h and then subjected to p62 immunoblotting. The results are presented as the means ± SEM from 3 independent experiments. The data were statistically analyzed by Student t-test or one-way ANOVA with Tukey’s post-hoc correction were used as indicated. *P < 0.05; **P < 0.01