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. 2021 Jun 1;11:11474. doi: 10.1038/s41598-021-90822-2

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© The Author(s) 2021

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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p62-mediated cleavage of TDP-43 is independent of caspases. NSC-34 cells were pre-treated with 20 μM caspase inhibitor, 5 μM calpain inhibitor or were left untreated for 1 h and then transfected with TDP-43-tdTomato and either EGFP or EGFP-p62. Cells were maintained in media containing the inhibitor for 48 h and then harvested for soluble and insoluble fractions. Western blot analyses for TDP-43 (anti-RFP), EGFP and α-tubulin were performed. TDP-43 bands were quantified using ImageJ and the amount of TDP-35-tdTomato fragment observed in the insoluble fraction of EGFP versus EGFP-p62 expressing cells was compared with SPSS.