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pH-independent entry and sequential endosomal sorting are major determinants of hepadnaviral infection in primary hepatocytes†
Article first published online: 29 AUG 2006
Copyright © 2006 American Association for the Study of Liver Diseases
Volume 44, Issue 3, pages 685–693, September 2006
How to Cite
Funk, A., Mhamdi, M., Hohenberg, H., Will, H. and Sirma, H. (2006), pH-independent entry and sequential endosomal sorting are major determinants of hepadnaviral infection in primary hepatocytes. Hepatology, 44: 685–693. doi: 10.1002/hep.21297
Potential conflict of interest: Nothing to report.
- Issue published online: 29 AUG 2006
- Article first published online: 29 AUG 2006
- Manuscript Accepted: 30 MAY 2006
- Manuscript Received: 3 APR 2006
- Freie und Hansestadt Hamburg
- Bundesministerium für Gesundheit (to the Heinrich-Pette-Institut)
- German Competence Network for Viral Hepatitis (Hep-Net)
- German Ministry of Education and Research (BMBF). Grant Number: TP13.1
Supplementary material for this article can be found on the H EPATOLOGY website ( http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html ).
|jws-hep.21297.fig1.tif||14573K||Supplementary Fig. 1.PtIns3PK signalling and Golgi apparatus are not involved in hepadnaviral transport.PDHs were treated with wortmannin to prevent PtIns3PK signalling (A) or BFA to disrupt Golgi (B) and subsequently infected with DHBV. Cells were harvested 3 d later and analysed by immunofluorescence staining as well as immunoblot for viral L protein. Wortm, wortmannin; BFA, brefeldin A; preT x , pre-treatment.|
|jws-hep.21297.fig2.tif||1980K||Supplementary Fig. 2.vATPase is required for a postentry step.PDHs were treated with BafA1 for 1 h at 37?C before inoculation with DHBV at 4?C for 2 h. Cells were immediately harvested by direct lysis after trypsin digestion or warmed to 37?C to allow internalization of bound viral particles. At the indicated time points, cells were harvested by limited protease digestion and analysed for intracellular viral DNA by PCR. +T, after trypsin digestion.|
|jws-hep.21297.fig3.tif||9358K||Supplementary Fig. 3.Disruption of endosomal transport stabilizes viral particles.PDHs were treated with the indicated substances for 1 h at 37?C. Then viral particles were allowed to attach to the cell surface and harvested (A). Parallel cultures were washed, supplemented with fresh drugs and shifted to 37?C for 6 h. Cells were then harvested and analysed by immunoblotting for viral L protein (B).|
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