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. 2022 Jun 30;13(6):575. doi: 10.1038/s41419-022-05030-1

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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 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. 6 — A Immunoblotting for α-synuclein in SH-SY5Y cells transfected with plasmids encoding vector alone or human Myc-caPAK4. B Quantification of α-synuclein intensity in blot A normalized to GAPDH. C In vitro ubiquitination of α-synuclein. D Immunoprecipitation (IP) and immunoblotting for the indicated E3 ligases. E, F Immunoprecipitation (IP) and immunoblotting. SH-SY5Y cell lysates were immunoprecipitated with anti-PAK4 or anti-NEDD4-1 (E)/anti-NEDD4-2 (F) antibodies followed by immunoblotting for PAK4, NEDD4-1, and GAPDH. G Immunoblotting for α-synuclein in SH-SY5Y cells following incubation with the indicated E3 ligase inhibitors. H Quantification of the α-synuclein signal in the blot E normalized to GAPDH. I Immunoblotting for α-synuclein in SH-SY5Y cells transfected with NEDD4-1 siRNAs. J Quantification of α-synuclein levels in the blot shown (I) normalized to GAPDH. The data are presented as the mean ± SEM. *P < 0.01. Two-way ANOVA (B, H, J) was used for statistical analysis followed by Tukey’s multiple comparisons test.