Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1

Daniel J. Klionsky, Amal Kamal Abdel-Aziz, Sara Abdelfatah, Mahmoud Abdellatif, Asghar Abdoli, Steffen Abel, Hagai Abeliovich, Marie H. Abildgaard, Yakubu Princely Abudu, Abraham Acevedo-Arozena, Iannis E. Adamopoulos, Khosrow Adeli, Timon E. Adolph, Annagrazia Adornetto, Elma Aflaki, Galila Agam, Anupam Agarwal, Bharat B. Aggarwal, Maria Agnello, Patrizia AgostinisJaved N. Agrewala, Alexander Agrotis, Patricia V. Aguilar, S. Tariq Ahmad, Zubair M. Ahmed, Ulises Ahumada-Castro, Sonja Aits, Shu Aizawa, Yunus Akkoc, Tonia Akoumianaki, Hafize Aysin Akpinar, Ahmed M. Al-Abd, Lina Al-Akra, Abeer Al-Gharaibeh, Moulay A. Alaoui-Jamali, Simon Alberti, Elísabet Alcocer-Gómez, Cristiano Alessandri, Muhammad Ali, M. Abdul Alim Al-Bari, Saeb Aliwaini, Javad Alizadeh, Eugènia Almacellas, Alexandru Almasan, Alicia Alonso, Guillermo D. Alonso, Nihal Altan-Bonnet, Dario C. Altieri, Élida M.C. Álvarez, Sara Alves, Cristine Alves da Costa, Mazen M. Alzaharna, Marialaura Amadio, Consuelo Amantini, Cristina Amaral, Susanna Ambrosio, Amal O. Amer, Veena Ammanathan, Zhenyi An, Stig U. Andersen, Shaida A. Andrabi, Magaiver Andrade-Silva, Allen M. Andres, Sabrina Angelini, David Ann, Uche C. Anozie, Mohammad Y. Ansari, Pedro Antas, Adam Antebi, Zuriñe Antón, Tahira Anwar, Lionel Apetoh, Nadezda Apostolova, Toshiyuki Araki, Yasuhiro Araki, Kohei Arasaki, Wagner L. Araújo, Jun Araya, Catherine Arden, Maria Angeles Arévalo, Sandro Arguelles, Esperanza Arias, Jyothi Arikkath, Hirokazu Arimoto, Aileen R. Ariosa, Darius Armstrong-James, Laetitia Arnauné-Pelloquin, Angeles Aroca, Daniela S. Arroyo, Ivica Arsov, Rubén Artero, Dalia Maria Lucia Asaro, Michael Aschner, Milad Ashrafizadeh, Osnat Ashur-Fabian, Atanas G. Atanasov, Alicia K. Au, Patrick Auberger, Holger W. Auner, Laure Aurelian, Riccardo Autelli, Laura Avagliano, Yenniffer Ávalos, Sanja Aveic, Célia Alexandra Aveleira, Tamar Avin-Wittenberg, Yucel Aydin, Scott Ayton, Srinivas Ayyadevara, Maria Azzopardi, Misuzu Baba, Jonathan M. Backer, Steven K. Backues, Dong Hun Bae, Ok Nam Bae, Soo Han Bae, Eric H. Baehrecke, Ahruem Baek, Seung Hoon Baek, Sung Hee Baek, Giacinto Bagetta, Agnieszka Bagniewska-Zadworna, Hua Bai, Jie Bai, Xiyuan Bai, Yidong Bai, Nandadulal Bairagi, Shounak Baksi, Teresa Balbi, Cosima T. Baldari, Walter Balduini, Andrea Ballabio, Maria Ballester, Salma Balazadeh, Rena Balzan, Rina Bandopadhyay, Sreeparna Banerjee, Sulagna Banerjee, Ágnes Bánréti, Yan Bao, Mauricio S. Baptista, Alessandra Baracca, Cristiana Barbati, Ariadna Bargiela, Daniela Barilà, Peter G. Barlow, Sami J. Barmada, Esther Barreiro, George E. Barreto, Jiri Bartek, Bonnie Bartel, Alberto Bartolome, Gaurav R. Barve, Suresh H. Basagoudanavar, Diane C. Bassham, Robert C. Bast, Alakananda Basu, Henri Batoko, Isabella Batten, Etienne E. Baulieu, Bradley L. Baumgarner, Jagadeesh Bayry, Rupert Beale, Isabelle Beau, Florian Beaumatin, Luiz R.G. Bechara, George R. Beck, Michael F. Beers, Jakob Begun, Christian Behrends, Georg M.N. Behrens, Roberto Bei, Eloy Bejarano, Shai Bel, Christian Behl, Amine Belaid, Naïma Belgareh-Touzé, Cristina Bellarosa, Francesca Belleudi, Melissa Belló Pérez, Raquel Bello-Morales, Jackeline Soares de Oliveira Beltran, Sebastián Beltran, Doris Mangiaracina Benbrook, Mykolas Bendorius, Bruno A. Benitez, Irene Benito-Cuesta, Julien Bensalem, Martin W. Berchtold, Sabina Berezowska, Daniele Bergamaschi, Matteo Bergami, Andreas Bergmann, Laura Berliocchi, Clarisse Berlioz-Torrent, Amélie Bernard, Lionel Berthoux, Cagri G. Besirli, Sebastien Besteiro, Virginie M. Betin, Rudi Beyaert, Jelena S. Bezbradica, Kiran Bhaskar, Ingrid Bhatia-Kissova, Resham Bhattacharya, Sujoy Bhattacharya, Shalmoli Bhattacharyya, Md Shenuarin Bhuiyan, Sujit Kumar Bhutia, Lanrong Bi, Xiaolin Bi, Trevor J. Biden, Krikor Bijian, Viktor A. Billes, Nadine Binart, Claudia Bincoletto, Asa B. Birgisdottir, Geir Bjorkoy, Gonzalo Blanco, Ana Blas-Garcia, Janusz Blasiak, Robert Blomgran, Klas Blomgren, Janice S. Blum, Emilio Boada-Romero, Mirta Boban, Kathleen Boesze-Battaglia, Philippe Boeuf, Barry Boland, Pascale Bomont, Paolo Bonaldo, Srinivasa Reddy Bonam, Laura Bonfili, Juan S. Bonifacino, Brian A. Boone, Martin D. Bootman, Matteo Bordi, Christoph Borner, Beat C. Bornhauser, Gautam Borthakur, Jürgen Bosch, Santanu Bose, Luis M. Botana, Juan Botas, Chantal M. Boulanger, Michael E. Boulton, Mathieu Bourdenx, Benjamin Bourgeois, Nollaig M. Bourke, Stephen L. Gregory

Research output: Contribution to journalReview articlepeer-review

203 Citations (Scopus)

Abstract

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
Pages (from-to)1-382
Number of pages382
JournalAutophagy
Volume17
Issue number1
Early online date8 Feb 2021
DOIs
Publication statusPublished - 2021

Keywords

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

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