Protocol for assessing translation in living Drosophila imaginal discs by O-propargyl-puromycin incorporation.

Translation is a fundamental process of cellular behavior. Here, we present a protocol for measuring translation in Drosophila epithelial tissues using O-propargyl-puromycin (OPP), a puromycin derivative. We detail steps for larval dissection, OPP incorporation, fixation, OPP labeling, immunostaining, and imaging. We also provide details of quantification analysis. Significantly, OPP addition to methionine-containing media enables polypeptide labeling in living cells. Here, we study wing imaginal discs, an excellent model system for investigating growth, proliferation, pattern formation, differentiation, and cell death. For complete details on the use and execution of this protocol, please refer to Lee et al. (2018), Ji et al. (2019), and Kiparaki et al. (2022).1,2,3. Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Citation

Marianthi Kiparaki, Nicholas E Baker. Protocol for assessing translation in living Drosophila imaginal discs by O-propargyl-puromycin incorporation. STAR protocols. 2023 Dec 15;4(4):102653

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PMID: 37862174

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