FIG 8.

The C-terminal region of CSFV NS5A is critical for the NS5A-GBP1 interaction. (A) Schematic representation of the protein domains of NS5A. Full-length NS5A and two deletion mutants were examined in this study. (B) Expression of full-length and truncated forms of NS5A. The indicated expression plasmids were transfected into HEK293T cells, and cell lysates were analyzed by Western blotting (WB) using antibodies against the Myc tag and GAPDH. (C and D) The C-terminal region of NS5A is required for its binding to GBP1 and GBP1(1-308). The full-length and truncated forms of NS5A were examined by co-IP analysis for interaction with GBP1 (C) or GBP1(1-308) (D). HEK293T cells were transfected with the indicated expression plasmids. Co-IP was performed using an anti-Flag monoclonal antibody (1:1,000). The precipitated proteins were analyzed by Western blotting using antibodies against the Myc and Flag tags.