Fig. 1.

Generation of CLDN1-deficient Huh-7.5 cells. (A) Immunoblotting analysis for GFP, CLDN1, or actin was performed on lysates from naïve Huh-7.5 cells or CLDN1 KO Huh-7.5 cell clones either not expressing a transgene (not transduced) or transduced with lentiviral particles to express GFP-CLDN1. Approximate molecular mass (kDa) marker positions are indicated to the left of each blot. (B) Above cells were infected with GLuc-expressing Jc1 in the presence of DMSO (vehicle, light gray columns) or the HCV polymerase inhibitor, 2′CMA (dark gray columns). Parallel VSVGpp infections were performed as a positive control (white columns). Luciferase values are expressed as RLU. Mean and SE of two independent experiments, each performed in triplicate, are shown. (C) Naïve Huh-7.5 cells were infected with supernatants from the HCV+DMSO infections shown in (B). (D) Relative titers of non-reporter Jc1 virus were determined by limiting dilution assay on naïve and CLDN1 KO clone 4 Huh-7.5 cells either not transduced or expressing GFP-CLDN1. Values represent means and SE of three independent virus preparations and are expressed as TCID50/mL. Limit of detection is indicated by a dashed line. Abbreviations: DMSO, dimethyl sulfoxide; ND, not detected; RLU, relative light units; TCID50/mL, 50% tissue culture infectious dose per milliliter.