Figure 2.
The substitutions I696T and I696N conferred AR4A resistance to J6/JFH1ΔHVR1. A, Huh7.5 cells were transfected with in vitro–transcribed RNA of the indicated recombinants harboring I696T, I696N, L726V or the combination I696N/L726V. Error bars represent standard errors of the mean (SEMs). Supernatants were collected at the indicated time points, and hepatitis C virus infectivity titers were determined. Lower level of quantification was 100 focus-forming units (FFU)/mL. B, First passages of the viruses were subjected to a dilution series of AR4A, from 50 µg/mL to 0.000005 µg/mL, and neutralization was assessed and analyzed as described in Figure 1, except we used 4-parameter curve fitting to obtain a sigmoidal dose-response curve. Error bars represent SEMs. Independent neutralization experiments yielded similar results.