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. 2017 Aug 23;7:9223. doi: 10.1038/s41598-017-10087-6

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Shows level of HCV replication in non-proliferative primary human hepatocytes and proliferative Huh-7.5 cell cultures. PHHs were infected with HCV-GFP virus (JFH-AM120) at a MOI of 0.1 by overnight incubation. The next day cells were incubated with fresh media with 10% human serum. (A) HCV replication in PHHs model was confirmed by the measurement of NS3 and NS5A-GFP fusion protein levels by Western blot. (B) Western blot analysis showed time dependent increase in the expression of NS3 protein in Huh-7.5 cells infected with HCV-GFP virus. (C) Expression of NS5A-GFP chimera protein with DAPI staining in Huh-7.5 cells infected with HCV-GFP virus over 12 days. (D) Quantification of NS5A-GFP expression by ImageJ software. (E) Flow cytometry analysis shows expression of GFP in infected Huh-7.5 cells at day 0, 9 and 12 days. Fluorescence images shown in panel C were taken at 40X magnification.