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. 2019 Aug 15;9:11878. doi: 10.1038/s41598-019-48047-x

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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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Mitophagy is impaired in sacsin KO cells. (Panels A–F) Autophagosome-lysosome fusion is disrupted in sacsin KO cells. Immunofluorescence images of LC3 (in green) and LAMP1 (in red) in WT and sacsin KO cells under normal conditions (A,D), FCCP treatment (B,E) and FCCP treatment followed by medium recovery (C,F). DAPI (in blue) was used as nuclear stain. Fusion between autophagosomes and lysosomes was reduced in sacsin KO cells even after medium recovery. Colocalization of LC3 with LAMP1 (in yellow) is indicated by arrows. (Panels G–N) Mitophagy activation is reduced in KO cells. Immunofluorescence images of the mitochondrial marker TOM20 (in green) and LC3 (in red) in WT and sacsin KO cells under normal conditions (G,L) FCCP treatment (H,M) and FCCP treatment followed by medium recovery (I,N). DAPI (in blue) was used as nuclear stain. Fusion between mitochondria and autophagosomes was reduced in sacsin KO cells even after medium recovery. Colocalization of LC3 with TOM20 (in yellow) is indicated by arrows. Scale bar = 10 µm. (Re = normal medium recovery).