Formation of the autophagosome is likely the most complex step of macroautophagy, and indeed it is the morphological and functional hallmark of this process; accordingly, it is critical to understand the mechanisms that govern this process, which is essential for the proper functioning of autophagy. This research activity aims to investigate the role of Atg8–PE deconjugation in autophagosome formation, which is a crucial step in the autophagic pathway. The findings of this study will provide valuable insights into the regulation of autophagosome formation and its implications for the development of novel therapeutic strategies for autophagy-related diseases.