FIG 3.

Interaction between heparin and HCV envelope proteins. Different volumes of lysates (200 μl, 100 μl, or 50 μl) obtained from cells infected with JFH1 or JFH1-ΔHVR1 were preincubated in the presence or absence of 500 μg/ml heparin for 1 h at 4°C and then precipitated with heparin-conjugated beads. Proteins were eluted with Laemmli buffer, and samples were separated on SDS-polyacrylamide gels. (A) HCV envelope proteins were detected by immunoblotting with A4 (anti-E1) and A11 (anti-E2) antibodies. (B) The quantities of E1 and E2 precipitated under each condition are presented. Results are expressed as the percentage of the total protein input. Data were analyzed by using Student's t test (*, P < 0.01). Mean values from three independent experiments are given. Error bars represent the standard deviations of the means. (C) HCV envelope proteins precipitated with heparin-conjugated beads from purified virus JFH1 or JFH1-ΔHVR1 lysates were detected by immunoblotting with A4 (anti-E1) and A11 (anti-E2) antibodies. (D) Purified JFH1 or JFH1-ΔHVR1 viruses were precipitated with heparin-coated beads, anti-apoE-coated protein G beads, or protein G beads. The amount of virus that bound to the beads was measured by quantitative RT-PCR. Mean values from three independent experiments are given. Error bars represent the standard deviations of the means.