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Updated: Mar 21, 2026

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells
Published on: May 16, 2017
Analysis of CRM1-Dependent Nuclear Export in Permeabilized Cells
Ralph H Kehlenbach1, Sarah A Port2
1Faculty of Medicine, Institute of Molecular Biology, University of Göttingen, Humboldtallee 23, 37073, Göttingen, Germany. rkehlen@gwdg.de.
Abstract:
Nuclear protein import and export assays in permeabilized cells have been instrumental for the identification of transport factors and for the molecular characterization of nucleocytoplasmic transport pathways. Our original assay to quantitatively analyze CRM1-dependent export was based on stably transfected cells expressing GFP-NFAT. We now present a simplified version of the assay using transiently transfected cells expressing GFP-NFAT or GFP-snurportin1 as a fluorescent export cargo and mCherry-emerin as a marker protein for transfected cells. CRM1- and Ran-dependent export is recapitulated in digitonin-permeabilized cells and quantified by flow cytometry. The assay should be applicable to other combinations of cargo and marker proteins.
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