Potential conflict of interest: Nothing to report.
Cell entry, efficient RNA replication, and production of infectious hepatitis C virus progeny in mouse liver-derived cells
Article first published online: 18 NOV 2013
© 2013 by the American Association for the Study of Liver Diseases
Volume 59, Issue 1, pages 78–88, January 2014
How to Cite
Frentzen, A., Anggakusuma, Gürlevik, E., Hueging, K., Knocke, S., Ginkel, C., Brown, R. J.P., Heim, M., Dill, M. T., Kröger, A., Kalinke, U., Kaderali, L., Kuehnel, F. and Pietschmann, T. (2014), Cell entry, efficient RNA replication, and production of infectious hepatitis C virus progeny in mouse liver-derived cells. Hepatology, 59: 78–88. doi: 10.1002/hep.26626
Supported by a grant from European Research Council ERC-2011-StG_281473-(VIRAFRONT) and by a grant from the Helmholtz Association SO-024 to T.P.
- Issue published online: 20 DEC 2013
- Article first published online: 18 NOV 2013
- Accepted manuscript online: 19 JUL 2013 05:35AM EST
- Manuscript Accepted: 3 JUL 2013
- Manuscript Received: 14 MAY 2013
Additional Supporting Information may be found in the online version of this article.
|hep26626-sup-0002-suppfig1.tif||14032K||Supporting Information Figure 1. MLT-MAVS-/- cells display characteristics of hepatocellular carcinoma (HCC) in vivo. C57BL/6 mice were subcutaneously implanted with 1x107 MLT-MAVS-/- cells and tumor sections were stained with (A) H&E for histopathological analysis or (B) with CK8, CK18, and CK19 specific antibodies for immunohistochemical characterization. As a control for the CK19 stain, a positive murine CCC sample with distinct structure was stained simultaneously for the purpose of comparison.|
|hep26626-sup-0003-suppfig2.tif||1127K||Supporting Information Figure 2. HCV Replication in MLT-MAVS-/-miR-122 derivatives is inhibited by mouse IFNα-1 and 2'CMA. Cell lines expressing ApoE and HCV entry receptors were transfected with Luc-Jc1 RNA followed by treatment with either 500 U/mL mIFNα-1 or 5 ug/mL 2'CMA 4 h post transfection. Luciferase activity was determined 48 h post transfection and depicted results are the means and standard deviations of at least two independent experiments.|
|hep26626-sup-0004-suppfig3.eps||2419K||Supporting Information Figure 3. Receptor expression of MLT-MAVS-/- cells and derivatives expressing human or mouse ApoE and HCV entry factors. Flow cytometry was used to determine the expression of endogenous or ectopically expressed human or mouse CD81 (A) or SCARB1 (B) using appropriate antibodies. (C) Expression of human or mouse OCLN and CLDN1 was determined by immunoblotting. Table S1 lists individual cell lines analyzed.|
|hep26626-sup-0005-suppfig4.tif||4474K||Supporting Information Figure 4. HCV RNA-replication in MLT-MAVS-/-miR-122 derived cell lines. Cell lines analyzed in Fig. 5 were transfected with a subgenomic HCV reporter replicon and replication was analyzed by luciferase assay at the indicated time points. Mean values +SD from at least three independent experiments are given.|
|hep26626-sup-0006-suppfig5.tif||2290K||Supporting Information Figure 5. Infection of MLT-MAVS-/- miR-122 derived cell lines with HCVpp and HCVTCP. (A) MLT-MAVS-/-miR-122 derived cell lines expressing either complete or minimal human entry factors were infected with HCVpp of GT1a and GT2a. 48 h post infection cells were lysed and luciferase activity was measured. (B) Cells in (A) were challenged with HCVTCP and 48 h post infection luciferase activity was determined. Mean values +SD of three independent experiments are shown.|
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