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. 2024 Jan 29;43(5):754–779. doi: 10.1038/s44318-024-00036-1

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© The Author(s) 2024

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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) NDP52 KO HeLa cells expressing Parkin, GFP alone, or GFP-tagged monobodies (MonoB) were treated with or without valinomycin (val) and bafilomycin (baf) for 1.5 h. The cells were immunostained with anti-OPTN and TOMM20 antibodies. Bars, 10 μm. (B) The cells in (A) were analyzed by immunoblotting (IB). The light blue and orange arrowheads indicate unmodified and ubiquitinated protein bands, respectively. (C) The levels of pTBK1 and pOPTN in (B) were quantified. The protein levels in cells without MonoB after 1 h val and baf treatment were set to 100. Error bars represent mean ± s.d. of three independent experiments. n.s. not significant, ***P < 0.001 by two-tailed Dunnett’s test. (D) Proposed model of TBK1 activation during PINK1/Parkin-mediated mitophagy. Source data are available online for this figure.