Methods to analyze SNARE-dependent vesicular fusion events that regulate autophagosome biogenesis.
Bottom Line: Autophagy is an important catabolic pathway that preserves cellular homeostasis.The formation of autophagosomes is a complex process requiring the reorganization of membranes from different compartments.These two steps are essential for the maturation of small single-membrane autophagic precursors containing ATG16L1 and mATG9 proteins into double-membrane autophagosomes.
Affiliation: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.Show MeSH
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Mentions: The colocalisation between ATG16L1 and mATG9 can also be measured in whole cells by classical immunofluorescence and confocal analysis. Loss of interaction between the ATG16L1 and mATG9 compartments was observed again in cells where VAMP3 was silenced, as well as in cells placed for 4 h at 18 °C, where the membrane traffic between the early endosomes and recycling endosomes was inhibited (Fig. 6).
Affiliation: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.