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

Marianthi Kiparaki; Nicholas E. Baker

doi:10.1016/j.xpro.2023.102653

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

Marianthi Kiparaki, Nicholas E. Baker

Genetics

Research output: Contribution to journal › Article › peer-review

Abstract

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

Original language	English (US)
Article number	102653
Journal	STAR Protocols
Volume	4
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2023.102653
State	Published - Dec 15 2023

Keywords

Developmental biology
Genetics
Model Organisms
Molecular/Chemical Probes

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2023.102653

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abstract = "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",

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

Abstract

Keywords

ASJC Scopus subject areas

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