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 method described above can be also be used to analyze the heterotypic fusion between ATG16L1 and mATG9 carrying vesicles. Here we show how this method can be used to analyze the heterotypic fusions between ATG16L1 and mATG9 vesicles and provide a control where the critical SNARE, VAMP3, was depleted. Silencing of VAMP3 led to a strong decrease of the colocalisation between the green and red vesicles (Fig. 5).
Affiliation: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.