Glycosylation profiling of HA and NA in mammalian VLPs.

<p>Mock-treated and deglycosylase-treated proteins of H5N1-VLPs were separated by SDS-PAGE, stained with Coomassie blue (A), and further analyzed by Western blotting using antibodies against H5 (B) and N1 (C). In parallel, mock-treated and deglycosylase-treated proteins of H3N2-VLPs were analyzed by Western blotting using the antibody against H3 (D). Untreated and deglycosylase-treated proteins of respective VLPs and virus are compared in panels B and D. Glycosylated HA1, HA2, NA, and H3 are denoted as HA1**, HA2**, NA**, and H3**, respectively. <b>#</b> represents glycosylated proteins harboring a residue moiety of complex-type glycans sensitive to PNGase F, but not Endo-H. <b>π</b> represents unknown posttranslational modifications on the NA of H5N1-VLPs.