FIG. 1.

Expression of recombinant NiV F and G glycoproteins. The NiV F and G glycoprotein ORFs were subcloned into the vaccinia virus promoter-driven expression vector pMC02 (8), and recombinant viruses were made (see Materials and Methods). HeLa cells were infected with NiV F- or G-encoding viruses and incubated 16 h at 37°C. Beginning at 6 h postinfection, cells were either labeled overnight with [³⁵S]methionine-cysteine for immunoprecipitation or cultured in medium alone for Western blotting. Lysates were prepared in a buffer containing Triton X-100 and clarified by centrifugation. Immunoprecipitation was performed with a rabbit anti-NiV or a rabbit anti-HeV antiserum followed by protein G-Sepharose. Western blotting was performed with a rabbit polyclonal antiserum against a synthetic F₂ peptide (see Materials and Methods). The metabolically labeled proteins were resolved by SDS-10% PAGE under reducing conditions and detected by fluorography; lysates for Western blotting were resolved by SDS-10% PAGE under reducing conditions and detected by chemiluminescence. (A and B) Immunoprecipitation; (C) Western blotting.