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. 2019 Nov 6;9:16150. doi: 10.1038/s41598-019-52534-6

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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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Truncating variants change the subcellular localization of parkin. (A) Localization of EGFP-tagged parkin clones in HEK293T cells. EGFP was directly visualized. Photographs were acquired using a Zeiss Axio Imager Z1 microscope. Bars, 10 μm. The percentage of cells with aggregates is also shown (graph). Data are presented as the mean ± SD of three independent experiments; ***p < 0.001 (ANOVA/Tukey). (B) Subcellular fractionation of HEK293T cells expressing parkin clones. Presence of parkin in each fraction was analyzed by immunoblotting with anti-EGFP antibody and normalized to the specific marker of each fraction. T, total fraction; Cp, cytoplasm fraction; Mb, membrane fraction; SN, soluble nuclear fraction; Ch, chromatin-bound nuclear fraction. Original blots are presented in Supplementary Fig. S4. Quantification data (graph) are presented as the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01 (one-way ANOVA/Tukey).