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. 2016 Oct 25;17(5):1357–1368. doi: 10.1016/j.celrep.2016.09.084

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© 2016 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Distinct Roles of NTCP in HDV/HBV and HCV Infections

(A) Flow cytometry analysis of HCV glycoprotein sE2 binding to Huh7.5.1 (Ctrl) and Huh7.5.1-NTCP (NTCP). Results are expressed as means ± SD percentage sE2 binding compared to control cells from three independent experiments (two performed in triplicate and one in quadruplicate; n = 10).

(B) sE2 binding inhibits HCVcc infection. Huh7.5.1 cells were treated with soluble sE2 and then infected with HCVcc (Luc-Jc1). Infection was assessed after 72 hr by measuring luciferase activity. Results are expressed as means ± SD percentage HCVcc infection from three independent experiments performed in triplicate (n = 9).

(C) Flow cytometry analysis of HCV entry factor expression in Huh7.5.1-NTCP cells. Results are expressed as percentage protein expression compared to parental Huh7.5.1 cells (set at 100%) from four independent experiments performed in duplicate (n = 8).

(D) Effect of HBV preS1-derived peptide on HDV, HCVpp (genotype 1b), and VSVpp entry into Huh7.5.1-NTCP cells. Huh7.5.1-NTCP cells were treated for 1 hr with preS1 or Ctrl peptide (200 nM). For HDV, cells were then infected with recombinant HDV for 7 days. Total RNA was purified and HDV RNA was detected by qRT-PCR. Results are expressed as percentage HDV RNA level compared to non-treated Huh7.5.1-NTCP cells from three independent experiments performed in triplicate (n = 9). For HCVpp (genotype 1b) and VSVpp, cells were infected with pseudoparticles and infection was assessed after 72 hr by luciferase activity. Results are expressed as means ± SD percentage viral entry from three independent experiments performed in quintuplicate (n = 15).

(E–G) Time-dependent inhibition of viral entry by preS1 treatment. Cells were treated with preS1 or Ctrl peptide for 24, 48, and 72 hr prior to infection with HCVpp (genotype 1b), VSVpp, or HDV. Infection was assessed as described above. (E) Results are expressed as means ± SD percentage HCVpp entry from five independent experiments performed in triplicate (n = 15). (F) Results are expressed as means ± SD percentage VSVpp entry from four independent experiments performed in triplicate (n = 12). (G) Results are expressed as means ± SD percentage HDV infection from three independent experiments performed in duplicate (n = 6).

(H) Treatment with preS1 inhibits HCVcc infection. Cells were treated with preS1 or Ctrl peptide for 72 hr prior to infection with HCVcc (Luc-Jc1). Infection was assessed after 72 hr by measuring luciferase activity. Results are expressed as means ± SD HCVcc infection (relative light unit, RLU) from three independent experiments performed in triplicate (n = 9).