FIG. 2.
In vitro and in vivo coimmunoprecipitation of various NS proteins and hVAP-33. (A) In vitro GST pull-down assay. In vitro-translated [35S]methionine-labeled NS5A was mixed with various GST fusion proteins. The precipitated NS5A protein was detected by autoradiography. (B and C) Various Flag-tagged and untagged constructs were cotransfected into Huh7 cells. At 48 h posttransfection, cell lysates were immunoprecipitated with anti-Flag-antibody cross-linked Sepharose-4B beads. An HCV patient serum was used to detect NS5A, NS5B, and NS4B by immunoblotting. Lanes 1, 3, 5, and 7 represent 10% of lysates prior to immunoprecipitation.