FIG. 3.

Protein μ2 is expressed from a recombinant baculovirus. The 8B M1 gene was subcloned into recombinant baculovirus by using the Bac-to-Bac baculovirus expression system (GIBCO BRL, Grand Island, N.Y.). Control GUS-expressing recombinant baculovirus was provided by the manufacturer. T. ni insect cells were infected with virus stock that had been passaged in Sf9 insect cells, and cell cultures were harvested at 48 h (B) or 72 h (A and B) postinfection (POSTINF), washed with phosphate-buffered saline supplemented with 1 mM phenylmethylsulfonyl fluoride, and lysed in radioimmunoprecipitation assay buffer. Lysate supernatants were resolved by electrophoresis on 10% Laemmli SDS–polyacrylamide gels. (A) Coomassie blue staining, duplicate samples. MW, molecular mass (kilodaltons) markers. (B) Western Blot analysis. For Western blot analysis, protein was transferred to Immobilon-P membrane with a semidry blotting system (Millipore, Bedford, Mass.). Detection by the ECL (enhanced chemiluminescence) system (Amersham Life Sciences, Arlington Heights, Ill.) was done according to the manufacturer’s protocol with hyperimmune rabbit antiserum as for Fig. 1 immunoprecipitations.