Work

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

Year: 2022

Type: article

Abstract: 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).

Source: STAR Protocols

DOI: 10.1016/j.xpro.2022.101813

Authors: Haochen Cui, Masood Sepehrimanesh, Casey A. Coutee, Masuma Akter, Md Abir Hosain, Baojin Ding

Institution: Louisiana State University Health Sciences Center Shreveport

Language: English

Topic: Nuclear Structure and Function

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,500

Funders: U.S. Department of Defense, National Institutes of Health, National Institute of Neurological Disorders and Stroke, LSU Health Shreveport