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Quantification of endogenous and exogenous protein expressions of Na,K-ATPase with super-resolution PALM/STORM imaging
KTH, School of Engineering Sciences (SCI), Applied Physics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Applied Physical Chemistry.
KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences (SCI), Applied Physics.ORCID iD: 0000-0003-0578-4003
2018 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 13, no 4, article id e0195825Article in journal (Refereed) Published
Abstract [en]

Transient transfection of fluorescent fusion proteins is a key enabling technology in fluorescent microscopy to spatio-temporally map cellular protein distributions. Transient transfection of proteins may however bypass normal regulation of expression, leading to overexpression artefacts like misallocations and excess amounts. In this study we investigate the use of STORM and PALM microscopy to quantitatively monitor endogenous and exogenous protein expression. Through incorporation of an N-terminal hemagglutinin epitope to a mMaple3 fused Na,K-ATPase (α1 isoform), we analyze the spatial and quantitative changes of plasma membrane Na,K-ATPase localization during competitive transient expression. Quantification of plasma membrane protein density revealed a time dependent increase of Na,K-ATPase, but no increase in size of protein clusters. Results show that after 41h transfection, the total plasma membrane density of Na,K-ATPase increased by 63% while the endogenous contribution was reduced by 16%. © 2018 Bernhem et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Place, publisher, year, edition, pages
Public Library of Science , 2018. Vol. 13, no 4, article id e0195825
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Physical Sciences
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URN: urn:nbn:se:kth:diva-227364DOI: 10.1371/journal.pone.0195825ISI: 000430802400044Scopus ID: 2-s2.0-85045925649OAI: oai:DiVA.org:kth-227364DiVA, id: diva2:1212861
Note

Export Date: 9 May 2018; Article; CODEN: POLNC; Correspondence Address: Brismar, H.; Science for Life Laboratory, Department of Applied Physics, Royal Institute of TechnologySweden; email: brismar@kth.se. QC 20180604

Available from: 2018-06-04 Created: 2018-06-04 Last updated: 2018-06-04Bibliographically approved

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Bernhem, KristofferBrismar, Hjalmar

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