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. 2022 Jan 10;12:351. doi: 10.1038/s41598-021-04131-9

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Establishment of a cell-based assay to evaluate αSyn aggregation. (A) Representative images of αSyn-EGFP aggregation in HeLa cells. HeLa cells were transfected with the indicated plasmids, and then treated with PBS, αSyn monomers, or PFF. (B) Quantification of the percentage of cells containing obvious αSyn-EGFP inclusion bodies in (A). Data are shown as the mean ± SEM of twelve independent wells (n = 12; **P < 0.01; two-way ANOVA with the Tukey test). (C) Representative immunocytochemistry images of PFF-treated HeLa cells. Cells were transfected with pCMV αSyn-EGFP, and then treated with αSyn PFF. Phosphorylated αSyn (pS129 αS) was stained using a specific antibody. (D,E) Quantification of the percentage of cells containing obvious αSyn-EGFP (D) and p129-αSyn (E) inclusion bodies. Data are shown as the mean ± SEM of four independent wells (n = 4; *P < 0.05; **P < 0.01; one-way ANOVA with the Dunnett test compared with the no αSyn PFF control).