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. 2017 Jul 19;7:5816. doi: 10.1038/s41598-017-04449-3

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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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66E2 does not inhibit HCV RdRp in vitro. (A) Coomassie stained SDS-PAGE depicting HCV RdRp lacking C-terminal 21 amino acids (3a NS5B Δ21) that was expressed in E. coli and purified to near homogeneity (lane 2). Lane 1, Molecular weight marker. (B) RNA synthesis in vitro by the 3a NS5B Δ21 in the presence of 66E2 and other known HCV inhibitors. The sequence of the template (LE21) used is given on the top. The amount of the 21-nt de novo initiated product synthesized was quantified and given as percent synthesis relative to the sample treated with only DMSO. The results presented are representative of three independent assays. (C) G418 resistant HCV-3a replicon expressing Huh7.5 cells were treated with varying concentrations of inhibitors. RNA replication is presented as % RLU. Reaction treated with DMSO was considered as 100%. The assays were performed in triplicates and results presented are representative of three independent assays. The % mean is shown above the bars and the error bars are standard deviations.