Issue 21, 2010

Kinetics of NF-κBnucleocytoplasmic transport probed by single-cell screening without imaging

Abstract

Transport of protein and RNA cargoes between the nucleus and cytoplasm (nucleocytoplasmic transport) is vital for a variety of cellular functions. The studies of kinetics involved in such processes have been hindered by the lack of quantitative tools for measurement of the nuclear and cytosolic fractions of an intracellular protein at the single cell level for a cell population. In this report, we describe using a novel method, microfluidic electroporative flow cytometry, to study kinetics of nucleocytoplasmic transport of an important transcription factor NF-κB. With data collected from single cells, we quantitatively characterize the population-averaged kinetic parameters such as the rate constants and apparent activation barrier for NF-κB transport. Our data demonstrate that NF-κB nucleocytoplasmic transport fits first-order kinetics very well and is a fairly reversible process governed by equilibrium thermodynamics.

Graphical abstract: Kinetics of NF-κB nucleocytoplasmic transport probed by single-cell screening without imaging

Supplementary files

Article information

Article type
Paper
Submitted
11 Jun 2010
Accepted
17 Aug 2010
First published
08 Sep 2010

Lab Chip, 2010,10, 2911-2916

Kinetics of NF-κB nucleocytoplasmic transport probed by single-cell screening without imaging

J. Wang, B. Fei, Y. Zhan, R. L. Geahlen and C. Lu, Lab Chip, 2010, 10, 2911 DOI: 10.1039/C0LC00094A

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