HCV NS5A interacts with Abi1 through the domain I of NS5A and regions I + II of Abi1.
A, schematic illustration of both wild type and mutant constructs of the NS5A expression plasmid. B, Abi1 interacts with domain I of NS5A. HEK293T cells were cotransfected with FLAG-tagged Abi1 and Myc-tagged NS5A expression plasmids. Total cell lysates harvested at 48 h after transfection were immunoprecipitated (IP) with an anti-FLAG antibody, and bound proteins were immunoblotted (IB) with an anti-Myc antibody. The 10% input proteins are shown. C, schematic illustration of both wild type and mutant Abi1 expression plasmids. D, NS5A interacts with regions I + II of Abi1. HEK293T cells were cotransfected with Myc-tagged NS5A and FLAG-tagged Abi1 expression plasmids. At 36 h after transfection, cell lysates were immunoprecipitated with an anti-Myc antibody, and bound proteins were immunoblotted with an anti-FLAG antibody. Protein expressions of Myc-tagged NS5A and FLAG-tagged Abi1 were verified by immunoblotting with an anti-Myc or anti-FLAG antibody using the same cell lysates. The 10% input proteins are shown. Experiments were carried out in duplicate.