Fig 1.

p100pop exhibits enhanced replication fitness in various human liver cell lines. (A) Schematic illustration of the genome organization of the parental virus Jc1FLAG(p7-nsGLuc2A) of p100pop and the Jc1 variants used in this study. Gray dots indicate number and approximate position of noncoding mutations. Light blue dots display coding mutations. The purple triangle reflects the approximate position of the J6/JFH1 junction within the intragenotypic Jc1 chimera. This illustration was generated using BioRender.com. (B) Frequency of adaptive coding mutations detected in the consensus sequence of p100pop. (C) Huh-7 cells were infected with 500 HCV genome equivalents (GE)/cell of different Jc1 variants. (D) Huh-7.5 cells were infected with 10 HCV GE/cell of Jc1-PS and p100pop. (E) Huh-7 cells were infected with 100 HCV GE/cell of Jc1-PS and p100pop. (F) HuH6 cells expressing hCLDN1 were infected with 1,000 HCV GE/cell of Jc1-PS and p100pop. (G) HepG2 cells expressing a HAHA-tagged version of hCD81 were infected with 1,100 HCV GE/cell of Jc1-PS and p100pop. (H) Huh-7.5 miR-122 knockout cells were infected with 140 HCV GE/cell of Jc1 and p100pop. (C–H) After a medium change at 4 h.p.i., cell culture supernatant was collected at 72 h.p.i. and used to determine viral particle production using a limiting dilution assay. Means and individual values of (C–F, H) n = 3 and (G) n = 4 biological independent experiments are shown. Statistical significance was calculated using two-tailed, unpaired t test with significance levels of P < 0.05 = *, P < 0.01 = **, P < 0.001 = ***, P < 0.0001 = ****. TCID₅₀, 50% tissue culture infectious dose. Dashed lines represent the limit of detection (LOD) of the limiting dilutions assay.