Abstract: Autophagy is an evolutionarily conserved degradation process in eukaryotes from yeast to mammalian cells, which degrades damaged proteins, organelles, cytosol and invaded microorganism to maintain homeostasis. Based on data reported in the literature and our own experience, we summarize some commonly used methods in autophagy research, such as LC3 turnover, GFP resistance to degradation, p62, mRFP-GFP-LC3 live imaging, electron microscope and flow cytometry. We also provide some experimental tips and details that people should pay attention to because these factors frequently cause experimental variations. We hope to provide some experimental guidance and references for researchers that are new to the field of autophagy. This will help people to get accurate and reproducible experimental results as well as valid explanations. 

Key words: autophagy, autophagic flux, LC3, autophagy inhibitor