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. 2020 Mar 4;10:4040. doi: 10.1038/s41598-020-61051-w

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© The Author(s) 2020

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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HSD17B12 KD induces dispersion and disruption of HCV replication sites. (A) Huh7.5 parental cells (MOCK shNT), infected with JFH-1 and transduced with lentiviruses expressing shNT (HCV shNT) or shHSD17B12 (HCV shHSD17B12) were stained with anti-dsRNA antibodies as makers of viral RNA replication sites and imaged by confocal microscopy. Results are representative of multiple field views. Scale bar = 10 µm (B) Huh7.5 cells transduced with lentiviruses expressing shNT or shHSD17B12 and transfected with JFH-1 DNA plasmids for four days are analyzed for RNA susceptibility to exogenous nucleases. Cells were treated with micrococcal nuclease, digitonin and/or NP40 as indicated by a+, and the relative abundance of HCV RNA (left panel) and actin mRNA (right panel) levels were determined by qRT-PCR analysis. RNA levels of cells treated with nuclease only are normalized to one. Values represent mean ± SD from the analysis of two experiments. Unpaired student t-test was used. P values < 0.05 (*) are indicated in comparison with shNT treatment.