Comparison of A/Shanghai/02/2013 and A/Netherlands/219/2003 HA.

(A) Alignment of A/Shanghai/02/2013 and A/Netherlands/219/2003 HA amino acid residues 130 to 310. Predicted antigenic sites A (pink), B (orange), C (purple), D (yellow), and E (blue) are highlighted. Glycosylation site N141 is marked in a square. (B) Overlapping of A/Netherlands/219/2003 HA (grey, PDB: 4DJ6) and A/Shanghai/02/2013 HA (green, PDB: 4LN6) crystal structures. Only monomers are shown. Predicted antigenic sites A (pink), B (orange), C (purple), D (yellow), and E (blue) are labeled. RBS: receptor binding site. (C) Western blot comparison of A/Shanghai/02/2013 and A/Netherlands/219/2003 HA treated with and without PNGase F. Wild type (WT) HA and its mutants were detected with goat antisera against A/Netherlands/219/2003 HA (Top panel). The p24 in HA-pseudoviruses, which served as loading controls, were detected with mouse mAb (183-H12-5C) against HIV-1 p24 Gag (Bottom panel). Data shown are representative of two independent experiments.