Kwon, Oh Yeun Kwon, Ick Chan Song, Hyun Kyu Jeon, Hyesung 2024-01-20T22:06:25Z 2024-01-20T22:06:25Z 2021-09-03 2008-12 0304-4165 https://pubs.kist.re.kr/handle/201004/132941 Background: The transcription factor NF-AT plays a key role in the activation of many early immune response genes and is regulated by subcellular localization. NF-AT translocates from the cytoplasm to the nucleus then returns in response to the intracellular calcium level. Methods: We have investigated NF-AT nucleocytoplasmic shuttling in real-time in living cells using NF-ATc1 tagged with the reversibly photoswitchable fluorescence protein, Dronpa. We monitored both nuclear import and export rate of Dronpa-tagged NF-AT in live cells upon stimulation with ionomycin plus calcium (I+Ca2+) or cyclosporin A (CsA). Results: The results show that NF-AT moved into the nucleus within 3-9 min after stimulation and moved back out into the cytoplasm within 15-50 min after CsA addition. In the absence of stimulation, NF-AT stayed in the cytoplasm as in the cells overexpressing GSK-3 beta, a calcineurin-opposing regulator. General Significance: This semi-quantitative imaging with constant fluorescence provides the basis to detect the real-time effect by several regulators on NF-AT family proteins. (C) 2008 Elsevier B.V. All rights reserved. English ELSEVIER SCIENCE BV HIPPOCAMPAL-NEURONS CYCLOSPORINE-A NUCLEAR CALCINEURIN Real-time imaging of NF-AT nucleocytoplasmic shuttling with a photoswitchable fluorescence protein in live cells Article 10.1016/j.bbagen.2008.08.003 1 BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v.1780, no.12, pp.1403 - 1407 BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS 1780 12 1403 1407 scie scopus 000260620800004 2-s2.0-52949107584 Biochemistry & Molecular Biology Biophysics Biochemistry & Molecular Biology Biophysics Article HIPPOCAMPAL-NEURONS CYCLOSPORINE-A NUCLEAR CALCINEURIN Real-time imaging Nucleocytoplasmic shuttling NF-AT Dronpa Calcineurin GSK-3 beta