Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system

Nucleocytoplasmic transport (NCT) plays critical roles in maintaining cellular homeostasis. Here, we present a protocol to measure NCT for both transcript and protein cargos in cultured cells. We first describe the fluorescent in situ hybridization (FISH) assay to measure the nuclear mRNA export. We...

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Autores principales: Cui, Haochen, Sepehrimanesh, Masood, Coutee, Casey A., Akter, Masuma, Hosain, Md Abir, Ding, Baojin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9641092/
https://www.ncbi.nlm.nih.gov/pubmed/36386872
http://dx.doi.org/10.1016/j.xpro.2022.101813
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author Cui, Haochen
Sepehrimanesh, Masood
Coutee, Casey A.
Akter, Masuma
Hosain, Md Abir
Ding, Baojin
author_facet Cui, Haochen
Sepehrimanesh, Masood
Coutee, Casey A.
Akter, Masuma
Hosain, Md Abir
Ding, Baojin
author_sort Cui, Haochen
collection PubMed
description Nucleocytoplasmic transport (NCT) plays critical roles in maintaining cellular homeostasis. Here, we present a protocol to measure NCT for both transcript and protein cargos in cultured cells. We first describe the fluorescent in situ hybridization (FISH) assay to measure the nuclear mRNA export. We then detail a dual reporter system to measure the protein NCT. This protocol also includes image analysis and data output using CellProfiler™. The combined approach can be used to unbiasedly analyze NCT activities in cultured cells. For complete details on the use and execution of this protocol, please refer to Ding et al. (2020, 2021).
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spelling pubmed-96410922022-11-15 Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system Cui, Haochen Sepehrimanesh, Masood Coutee, Casey A. Akter, Masuma Hosain, Md Abir Ding, Baojin STAR Protoc Protocol Nucleocytoplasmic transport (NCT) plays critical roles in maintaining cellular homeostasis. Here, we present a protocol to measure NCT for both transcript and protein cargos in cultured cells. We first describe the fluorescent in situ hybridization (FISH) assay to measure the nuclear mRNA export. We then detail a dual reporter system to measure the protein NCT. This protocol also includes image analysis and data output using CellProfiler™. The combined approach can be used to unbiasedly analyze NCT activities in cultured cells. For complete details on the use and execution of this protocol, please refer to Ding et al. (2020, 2021). Elsevier 2022-11-03 /pmc/articles/PMC9641092/ /pubmed/36386872 http://dx.doi.org/10.1016/j.xpro.2022.101813 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Cui, Haochen
Sepehrimanesh, Masood
Coutee, Casey A.
Akter, Masuma
Hosain, Md Abir
Ding, Baojin
Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
title Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
title_full Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
title_fullStr Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
title_full_unstemmed Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
title_short Protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
title_sort protocol to image and quantify nucleocytoplasmic transport in cultured cells using fluorescent in situ hybridization and a dual reporter system
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9641092/
https://www.ncbi.nlm.nih.gov/pubmed/36386872
http://dx.doi.org/10.1016/j.xpro.2022.101813
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