Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperReviewResearchpeer-review

  • Stig U Andersen
  • Tapas Chakraborty, Ehime Univ, Ehime University, Ctr Marine Environm Studies, Department of Aquatic Life Sciences, Ainan
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  • Wei Chen, Duke University & Duke University Medical Center, Durham Law School, Durham University , NC State, NC, USA
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  • Xin-Ming Chen, Sydney Medical School - Central, University of Sydney, Edward Ford Building A27, The University of Sydney, NSW 2006, Sydney, AUSTRALIA., Sydney Medical School, University of Sydney, Australia., Department of Renal Medicine, Pain Management Research Institute, Kolling Institute, Northern Sydney Local Health District, St Leonards, Sydney, NSW Dept Primary Ind, NSW Department of Primary Industries
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  • Yan Chen, Shanghai Institute of Nutrition and Health, Univ Chinese Acad Sci, University of Chinese Academy of Sciences, CAS, Chinese Academy of Sciences
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  • Malene Hansen, Sanford Burnham Prebys Medical Discovery Institute, United Nations Development Program, Aging and Regeneration, 1] Department of Psychiatry, University of California, San Diego, La Jolla, CA, USA [2] Multimodal Imaging Laboratory, University of California at San Diego, La Jolla, CA, USA [3] Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA [4] Department of Radiology, University of California, San Diego, La Jolla, CA, USA., St. Mary's College of CA Moraga, CA, US
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  • Kun Huang, Huazhong University of Science and Technology, X Luo, Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China., Department of Pharmacy Practice and Clinical Pharmacy, School of Pharmacy, University of Ghana, Legon, Ghana., Wuhan Asia Heart Hospital, Wuhan Shi, China., Hubei University
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  • Christian A Hubner, Institute of General Practice and Family Medicine, Jena University Hospital, University of Jena, Jena, Germany., Thuringia
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  • Thomas E Jensen, University of Copenhagen
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  • Niels Jessen
  • Stefan H E Kaufmann, Department of Molecular Biology, Max Planck Institute for Infection Biology, BerlinDepartment of Molecular Biology, Max Planck Institute for Infection Biology, Berlin, Berlin-Brandenburg Center for Regenerative Therapies, Charité, University Medicine Berlin, Campus Virchow Klinikum, Berlin, Germany; Department of Cardiology, Charité, University Medicine Berlin, Campus Virchow Klinikum, Berlin; DZHK (German Centre for Cardiovascular Research), partner site Berlin, Berlin, Germany.
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  • Joo-Yong Lee, Chungnam Natl Univ, Chungnam National University, Dept Astron & Space Sci, Graduate School of Analytical Science and Technology (GRAST)
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  • Bin Li, College of Life Science and Technology, Beijing University of Chemical Technology, Jinan Univ, Jinan University, Dept Ecol, Guangzhou Med Univ, Guangzhou Medical University, Affiliated Hosp 5
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  • Jing Li, Virginia Commonwealth University, Virginia, Kyoto-McGill International Collaborative School in Genomic Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan., University of Virginia, Charlottesville, VA, Portland VA Med Ctr, Portland VA Medical Center, US Department of Veteran Affairs
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  • Jiong Li, Sichuan Agr Univ, Sichuan Agricultural University, Triticeae Res Inst, West China Hospital of Sichuan University, State key laboratory of biotherapy, Chengdu Univ Technol, Chengdu University of Technology, Coll Environm & Ecol
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  • Qing Li, Hong Kong Institute of Diabetes and Obesity, The Chinese University of Hong Kong, Hong Kong, China., Department of Anaesthesia and Intensive Care Medicine, Cardiothoracic Anaesthesia and Intensive Care, University Hospital of Pisa, Pisa, Italy.
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  • Yuan Li, Department of Pediatrics, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA; Department of Computer Science, Stanford University, Stanford, CA 94305, USA., St. Mary's College of CA Moraga, CA, US
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  • Paula Lindner
  • Wei Liu, The First Affiliated Hospital, Institute of Translational Medicine, School of Medicine, Zhejiang University, Hangzhou 310058, China., Department of Biochemistry and Department of Cardiology of the Second Affiliated Hospital, Hangzhou ivy dental clinic Co., Limited , Hangzhou , China.
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  • Anders H Lund, University of Copenhagen
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  • Sara Martin, Ist Europeo Oncol IEO, IRCCS European Institute of Oncology (IEO), Div Canc Prevent & Genet, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan and University of Milan, Milan, Italy., Univ Milano Bicocca, University of Milano-Bicocca, Dipartimento Sci Mat, Division of Early Drug Development, Department of Oncology and Hemato-Oncology, European Institute of Oncology, University of Milano, Milano, Italy.
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  • Trine H Mogensen
  • Andreas Buch Møller
  • Luca M Neri, ENT Clinic, University of Ferrara, Ferrara, Italy.
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  • Morten Petersen, University of Copenhagen
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  • Martin Post, The Hospital for Sick Children, Graduate Program in Translational Medicine, Toronto Western Hosp, University of Toronto, University Health Network Toronto
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  • Chen Wang, China Pharmaceutical University, School of Life Science and Technology, ShanghaiTech University, Shanghai, China., Nanjing Medical University, Nanjing, China, Jiangsu Univ, Jiangsu University, Inst Adv Mat
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  • Hao Wang, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, Beijing 100190, China, CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience and Technology of China, Beijing 100190, China., National Center for Nanoscience and Technology (NCNST), Zhongguancun College
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  • Qing Wang
  • Qing Wang
  • Qing Wang
  • Yan Wang, Tsinghua Univ, Tsinghua University, Tsinghua Ctr Astrophys, Faculty of Health and Life Sciences, School of Life Sciences, Coventry University, Coventry, United Kingdom.
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  • Yu Wang, College of Agriculture, Nanjing Agricultural University, Nanjing, China, College of Horticulture, Nanjing Agricultural University
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  • Zhe Wang, Air Force Medical University, Xijing Hospital and School of Basic Medicine, Laboratory of Angiogenesis and Vascular Metabolism, VIB Center for Cancer Biology (CCB), VIB, 3000 Leuven, Belgium; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 51006, Guangdong, P.R. China; Laboratory of Angiogenesis and Vascular Metabolism, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, 3000 Leuven, Belgium., Xi'an Jiaotong University, Xi'an, Shaanxi
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  • Min Wu, University of North Dakota School of Medicine and Health Sciences, Grand Forks, North Dakota, USA., Division of Infection, Immunity and Respiratory Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, UK., Department of Biomedical Sciences, Energy and Environmental Research Center, University of North Dakota, Grand Forks, The 22.nd Nordic Congress on Psychology
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  • Yao-Wen Wu, Lund University, Malmö, Sweden; Umeå University, Umeå, Sweden., Umeå Centre for Microbial Research (UCMR)
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  • Hui Yang, Capital Medical University, School of Basic Medical Sceinces
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  • Ling Yang, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, Iowa., Fraternal Order of Eagles Diabetes Research Center, Department of Anatomy and Cell Biology, Western University, Ontario, Canada., Department of Psychiatry, University of Iowa, Iowa City, Iowa, USA., University of Northern Iowa, Cedar Falls, IA
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  • Qian Yang
  • Hong Zhang, Peking University First Hospital, Renal Division, Ghent University Hospital, Ghent, Belgium., Xi Cheng District
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  • Hong Zhang, Univ Chinese Acad Sci, University of Chinese Academy of Sciences, CAS, Chinese Academy of Sciences, National Laboratory of Biomacromolecules
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  • Lin Zhang, Hong Kong Institute of Diabetes and Obesity, The Chinese University of Hong Kong, Hong Kong, China., Department of Medicine and Therapeutics and Department of Anaesthesia and Intensive Care
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  • Lu Zhang, Henan University of Technology, College of Bioengineering, Chongqing University, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan,China
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  • Peng Zhang, Department of Chemistry, University of Utah, Utah., Huntsman Cancer Institute and Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah; Utah Center for Advanced Imaging Research, Salt Lake City, Utah., UT Health MD Anderson
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  • Xiaoyan Zhang, Department of Molecular, Cellular, and Developmental Biology, Yale University, Univ Michigan, University of Michigan, University of Michigan System, Pediat Genet, VA Ann Arbor Healthcare System and University of Michigan, Ann Arbor, Michigan., DGSA MI, Department of Surgery, School of Medicine, University of Michigan, Ann Arbor, Michigan 48109.
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  • Yang Zhang, Univ Houston, University of Houston, University of Houston System, Dept Earth & Atmospher Sci, Guangdong College of Pharmacy, Department of Pharmacological & Pharmaceutical Sciences, Ansh Labs, Webster, TX, USA
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  • Yuan-Yuan Zhang, Sichuan Agr Univ, Sichuan Agricultural University, Triticeae Res Inst, West China School of Basic Medical Sciences & Forensic Medicine, Chengdu Univ Technol, Chengdu University of Technology, Coll Environm & Ecol
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  • Zhe Zhang, East China Normal University, Key Laboratory of Adolescent Health Assessment and Exercise Intervention of Ministry of Education, Biomedical Instrument Institute, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China; Shanghai Med-X Engineering Research Center, Shanghai Jiao Tong University, Shanghai, China.
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  • Zili Zhang, Nanjing University of Chinese Medicine
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  • Hao Zhou, Fifth Medical Center of the General Hospital of the Chinese People's Liberation Army
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  • Wei-Xing Zong, Rutgers State Univ, Rutgers State University New Brunswick, Dept Genet, Chemical Biology Laboratory, Department of Biotechnology, Indian Institute of Technology, Madras (IITM), Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ, United States., RUTCOR, Rutgers University, Piscataway, NJ, Princeton University, Princeton, NJ

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

Original languageEnglish
JournalAutophagy
Volume17
Issue1
Pages (from-to)1-382
Number of pages382
ISSN1554-8627
DOIs
Publication statusPublished - Feb 2021

    Research areas

  • Autophagosome, LC3, cancer, flux, lysosome, macroautophagy, neurodegeneration, phagophore, stress, vacuole

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