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© Research
Publication : Methods in molecular biology (Clifton, N.J.)

Antibody uptake assay and in vivo imaging to study intracellular trafficking of Notch and Delta in Drosophila

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Methods in molecular biology (Clifton, N.J.) - 01 Jan 2014

Couturier L, Schweisguth F

Link to Pubmed [PMID] – 25053482

Methods Mol. Biol. 2014;1187:79-86

Notch signaling depends on regulated intracellular trafficking of the receptor and its ligands (Kopan and Ilagan, Cell 137:216-233, 2009; Le Borgne et al., Development 132:1751-1762, 2005). Here we describe two methods to study the intracellular trafficking of Notch and Delta in Drosophila. First, an ex vivo antibody uptake assay is used to monitor endocytosis of Notch and Delta by living cells in dissected explants (Le Borgne and Schweisguth, Dev Cell 5:139-148, 2003). Second, real-time imaging of fluorescent proteins that are expressed at physiological levels is used to study trafficking of Notch in living flies (Venken et al., Science 314:1747-1751, 2006; Couturier et al., Nat Cell Biol 14, 131-139, 2012).