Fig 2: Lipid-lowering drugs affect HCV infection of lipoprotein receptor-deficient cell lines. Drug-induced cell cytotoxicity of lipoprotein receptor-deficient cell lines (A). Huh-7.5 derived SR-B1 KO #3 (light orange bars). LDLr KO #1 (grey bars) and DKO #1 (light blue bars) were treated with 1 µM simvastatin, 5 µM fenofibrate or 1 µM alirocumab for 96 h. Cell viability was determined by MTT Assay. (B) HCVcc infection of KO cells (B). SR-B1 KO #3, LDLr KO #1, and DKO #1 were treated with indicated drugs 24 h before infection with an FLuc reporter GT2a HCV virus (Fluc-Jc1) in the presence of the compounds or solvent control. 4 h later, virus was removed and drugs applied for extra 72 h before cell lysis and FLuc measurement. FLuc activities were normalized to DMSO control. Graphs represent 3 independent experiments performed in triplicates. Bars show the average and error bars in SD. Black dots plot individual values. Multiple comparison two-way ANOVA with a post hoc Dunnett correction was performed for statistical analysis (a = 0.05, DF = 40). * p = 0.05; ** p = 0.01; **** p = 0.0001.

Fig 3: Lipoprotein receptor SR-B1 and LDLr do not affect HCV replication, SR-B1 facilitates GP-driven cell entry, and both redundantly participate in HCV infection. HCV GP-mediated entry in lipoprotein-deficient cells (A). Huh-7.5 parental (black bars), SR-B1 (light orange bars), LDLr (grey bars), and DKOs (light blue bars) were transduced with NLuc-encoding GT1a H77 or GT2a J6 HCVpp, negative (NoEnvpp), and positive (MLVpp) controls for 6 h. Six hpt, lentiviral particles were removed and reporter expression measured 72 h after. Raw relative light units (RLU) values were normalized to MLVpp values of the same cell line to account for intercellular variation. NoEnvpp normalized values were subtracted to represent HCV GP-specific entry. HCV subgenomic replication in Huh-7.5 and lipoprotein receptor-deficient cells (B). Cells were transfected with a HCV GT2a replication-competent (WT) or deficient (?GDD) subgenomic genomes tagged with FLuc. RLU values were normalized to 4 h post-electroporation and to MTT values to account for initial RNA delivery and cell growth variabilities between cell lines. Graph represents the average of three independent experiments performed in triplicate +/- sem. HCVcc infection of lipoprotein-deficient and parental cells (C). Indicated cell lines were infected untagged GT2a Jc1 HCVcc. Virus inoculum was removed hpi. Twenty-four hpi cells were lysed and Intracellular HCV RNA copies determined with an X-tail based absolute RT-qPCR. Data are shown as violin plots with median (black line) and individual values (black dots). Data distribution is symbolized by the plots’ shape. A two-way ANOVA with Dunnett post hoc (a = 0.05, DF = 10) was used for statistical analysis of HCV entry and replication assays and a one-way ANOVA with Sidak’s post hoc for HCVcc infection assay (a = 0.05, DF = 25). p-value was calculated and shown as: * p = 0.05; ** p = 0.01; *** p = 0.001; **** p = 0.0001.