Аннотация:In 2008, we published the first set of guidelines for standardizing research in autophagy. Since2545 then, this topic has received increasing attention, and many scientists have entered the field. Ourknowledge base and relevant new technologies have also been expanding. Thus, it is importantto formulate on a regular basis updated guidelines for monitoring autophagy in different organisms.Despite numerous reviews, there continues to be confusion regarding acceptable methodsto evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines2550 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 onthese processes. These guidelines are not meant to be a dogmatic set of rules, because theappropriateness of any assay largely depends on the question being asked and the system beingused. Moreover, no individual assay is perfect for every situation, calling for the use of multiple2555 techniques to properly monitor autophagy in each experimental setting. Finally, several corecomponents of the autophagy machinery have been implicated in distinct autophagic processes(canonical and noncanonical autophagy), implying that genetic approaches to block autophagyshould rely on targeting two or more autophagy-related genes that ideally participate in distinctsteps of the pathway. Along similar lines, because multiple proteins involved in autophagy also2560 regulate other cellular pathways including apoptosis, not all of them can be used as a specificmarker for bona fide autophagic responses. Here, we critically discuss current methods of assessingautophagy and the information they can, or cannot, provide. Our ultimate goal is toencourage intellectual and technical innovation in the field.