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. 2019 Feb 12;9:1847. doi: 10.1038/s41598-019-38526-6

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© The Author(s) 2019

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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Experimental design and cytotoxicity studies. (a) Structure of PCR products encoding sgRNAs. (b) Experimental design. HepG2-1.1merHBV or HepG2-1.5merHBV cells were lipofected with a Cas9-encoding plasmid and a PCR product encoding an sgRNA, and were treated with the small molecules B02, NU7026, 3-aza, L755, or DMSO simultaneously. Untransfected cells were negatively selected using blasticidin. (c) SgRNAs (Sp1-Sp3) used in this study, targeting functional regions of the HBV genome. Effects of small molecules on cell viability in (d) HepG2-1.1merHBV or (e) HepG2-1.5merHBV cell lines treated with small molecules for 72 h and observed for the designated period of time. Effects of small molecules on (f) HepG2-1.1merHBV cells or (g) HepG2-1.5merHBV cells 3 d post transfection with HBV-targeting CRISPR/Cas9 and concomitant treatment with small molecules. Asterisks indicate statistically significant differences. *p < 0.05.