Huh-7 and Huh-7.5 cells were cultured at 37°C, 5% CO2 in Dulbecco’s Modified Eagle Medium (DMEM, Invitrogen) containing 10% fetal bovine serum (FBS) and 0.1 mM nonessential amino acids (NEAA) (complete media), unless otherwise noted. For time-lapse imaging, cells were maintained in CO2-independent media (Invitrogen) containing 10% FBS, 0.1 mM NEAA, 1 mM sodium pyruvate and 2 mM L-glutamine (imaging media). Huh-7.5 cell lines harboring selectable subgenomic replicons8 (link) were grown in complete media containing 0.5 mg/ml G418. Huh-7.5 cells stably expressing the pLenti-3’-U6-EC-EP7 vector encoding an shRNA against CD81 (nt 268-288, cDNA numbering) or a predicted non-targeting sequence (IRR) have been previously described19 (link) and were grown in complete media containing 6 μg/ml blasticidin. MPCC cultures were generated as previously described23 (link) and maintained in high glucose DMEM, 10% FBS, 0.5 U/ml insulin, 7 ng/ml glucagon, 7.5 ug/ml hydrocortisone and 1% penicillin-streptomycin. For HCV inhibition, culture media was supplemented with 0.2% DMSO, 14 mM 2’CMA, 24 mM VX-950, or 1000 U/ml IFN-β (Peprotech). For neutralization experiments, HCVcc infections were performed in the presence 10 μg/ml antibody directed against CD81 (BD Biosciences) or an isotype control (IgG1κ, BD Biosciences).