| Literature DB >> 31250291 |
Marco Gualtieri1, Thomas Vaccari2.
Abstract
Mosaic analysis in Drosophila represents a convenient entry point for studying the role of ESCRT (Endosomal Sorting Complex Required for Transport) genes in multiple cell processes crucial for organ development and homeostasis. Here, we describe the procedure to generate populations of ESCRT-mutant cells within Drosophila larval epithelial organs and to study them in whole-mount preparations using confocal microscopy. The use of antibodies directed to endocytic cargoes, vesicular trafficking, cell proliferation, death, and polarity markers allows one to investigate the consequences of loss of ESCRT activity at the subcellular and tissue level. The protocols described here can be used in fixed tissue as well as in unfixed tissue using endocytic uptake assays.Entities:
Keywords: Confocal microscopy; Drosophila melanogaster; ESCRT function; Endocytosis; Imaginal discs; Whole-mount immunohistochemistry
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Year: 2019 PMID： 31250291 DOI： 10.1007/978-1-4939-9492-2_2
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745