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. 2018 Aug 24;12:262. doi: 10.3389/fncel.2018.00262

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Copyright © 2018 Zhu, Qiao, Chen, Sun, Hui, Lian, Xie, Ding, Meng, Zhu and Qiu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The level of SUMOylated α-syn in METH-treated cells increases when UBC9 is overexpressed. SH-SY5Y cells (A–C,A1–C1) and primary cultured neurons (D,E,D1,E1) were transfected with pcDNA3.1-NC or pcDNA3.1-UBC9 and LV-NC or LV-UBC9, respectively. Cells were treated with or without METH. Then, the cells were immunoprecipitated with an anti-SUMO-1 antibody, followed by western blot with an anti-α-syn antibody. Cells immunoprecipitated with IgG were used as a negative control. Western blot (A–E) and quantitative analyses (A1–E1) were performed to evaluate the efficiency of UBC9 overexpression, the expression of SUMO-1, and the expression, aggregation and SUMOylation of α-syn. β-Actin was used as a loading control. *p < 0.05 compared with the NC group. **p < 0.05 compared with the LV-UBC group. ^#p < 0.05 compared with the METH-treated NC group. The data were analyzed using one-way ANOVA followed by LSD post hoc analyses.