Journal
MICROBIAL PATHOGENESIS
Volume 51, Issue 3, Pages 225-229Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.micpath.2011.04.011
Keywords
Dengue; Endothelial cells; Envelope; NS1; Subcellular localization; Viral proteins
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Funding
- National Center for Genetic Engineering and Biotechnology (BIOTEC)
- Office of the Higher Education Commission
- Mahidol University under the National Research Universities Initiative
- Thailand Research Fund [RTA5380005]
- Siriraj Graduate Thesis Scholarship
- Faculty of Medicine Siriraj Hospital
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We performed subcellular fractionation of human endothelial cells (EA.hy926) after a challenge with dengue virus serotype-2 (DEN-2) at multiplicity of infection (MOI) of 10 for 2 h. The cells were fractionated into four fractions including cytosol, membrane/organelle, nucleus and cytoskeleton, and their purity was nicely confirmed by immunofluorescence staining of markers for individual subcellular compartments. Western blot analysis of dengue envelope (E) and non-structural 1 (NS1) proteins showed that E was present only in membrane/organelle fraction, whereas NS1 was present in membrane/organelle, nucleus, and cytoskeleton fractions. Time-course study showed that E was present immediately after the challenge, whereas NS1 was present after 12 h of the challenge. Our data may provide a new roadmap for studying mechanisms of replication of dengue virus and its interactions with host cells. (C) 2011 Elsevier Ltd. All rights reserved.
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