FIG. 4.

The 35K ORF 3 protein is phosphorylated. Sf9 cells were infected with wild-type baculovirus (WT) or the ORF 2, ORFs 2+3, ORF 3, or ORF 3HA baculovirus recombinants. Cells were harvested at 65 h p.i. in M-Per buffer plus protease inhibitors. An aliquot of the clarified lysate was removed for analysis (lanes L). The remaining sample was adjusted to 1× CIAP buffer and incubated in the absence (−) or presence (+) of CIAP (20 U). Western blot analysis was performed using the ORF 3 C-terminal peptide antiserum (1:500) (A) or the anti-rNV serum (1:1,000) (B). rNV 2+3 VLPs were included as a positive control for Western blot analysis.