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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

Bile Acid Uptake Enhances HCV Infection by Decreasing ISG Expression

(A) The impact of bile acid and preS1 treatment on IFITM3 expression. Huh7.5.1-NTCP cells were treated with the bile acid (BA) sodium taurocholate (100 μM) or with 200 nM preS1 for 72 hr. IFITM3 expression was then quantified by qRT-PCR. Results are expressed as means ± SD percentage IFITM3 expression compared to untreated (Ctrl) Huh7.5.1-NTCP cells (set at 100%) from three independent experiments performed in duplicate (n = 6).

(B) The impact of bile acid on HCVcc infection. Huh7.5.1 and Huh7.5.1-NTCP cells were treated 0, 25, or 100 μM sodium taurocholate for 72 hr and then infected with HCVcc (Luc-Jc1). Results are expressed as means ± SD percentage HCVcc infection compared to untreated (Ctrl) Huh7.5.1 and Huh7.5.1-NTCP cells (both set at 100%) from three independent experiments performed in triplicate (n = 9).

(C and D) Effect of preS1-mediated inhibition of bile acid uptake on IFITM3 protein expression and HCVcc infection. Huh7.5.1 and Huh7.5.1-NTCP cells were treated with sodium taurocholate (100 μM) in the presence or absence of preS1 peptide for 72 hr. (C) IFITM3 protein expression was assessed by western blot. One experiment is shown. (D) Cells were then infected by HCVcc (Jc1) for 3 days. Results are expressed as means ± SD percentage HCVcc infection compared to Huh7.5.1 and Huh7.5.1-NTCP cells treated with control peptide (both set at 100%) from three independent experiments performed in triplicate (n = 9).

(E and F) Silencing of IFITM3 expression increases HCV infection in a bile acid-dependent manner. Huh7.5.1-NTCP cells were transfected with siRNA control (siCtrl) or siRNA targeting IFITM3 (siIFITM3) and then treated for 72 hr in the absence or presence of BA (100 μM) in the presence of either preS1 or Ctrl peptide (200 nM). (E) Then 3 days after transfection, silencing efficacy was assessed by measuring expression of IFITM3 mRNA by qRT-PCR. Results are expressed as means ± SD percentage IFITM expression from four independent experiments performed in triplicate (n = 12). (F) Cells were 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 compared to cells treated with siCtrl (set at 100% for each condition) from four independent experiments performed in triplicate (n = 12).