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. 2021 Sep 27;11:19125. doi: 10.1038/s41598-021-98243-x

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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The inhibition of HBV attachment and internalization by CCM was depending on NTCP level. NTCP expression levels in HeLa, HepG2, Huh7, HepaRG, imHC, d-HepaRG and d-imHC was determined using qRT-PCR (A). The levels of cell bound HBV DNA that represented immediate HBV attachment was measured using real-time qPCR (B). A direct correlation between NTCP levels and the amount of bound HBV DNA was observed using Pearson's r (C). NTCP density in d-imHC was evaluated by flow cytometry (D). CCM inhibited HBV attachment in dose-dependent manner (E). CsA and HP served as known entry inhibitors. CCM inhibited HBV internalization in d-imHC (F). Data were presented as mean ± SD. *, **, *** and **** represented statistical difference with a p-value < 0.05, < 0.01, < 0.001 and < 0.0001 respectively.