Multiple sequence alignment of HA in the vicinity of the proteolytic cleavage site. (A) Sequence alignment of H1 HA. A total of 3,425 H1 HA sequences from the NCBI Influenza Virus Resource (http://www.ncbi.nlm.nih.gov/genomes/FLU/FLU.html) were aligned using Clustal W, and the aligned sequences of representative viruses at the HA cleavage site are shown, along with the consensus sequence. Dots indicate identical residues, and the P2-P1′ positions of the cleavage site and the location of the fusion peptide are shown. The representative viruses shown are the following: A/South Carolina/1/1918 (accession number AAD17229), A/PR/8/1934 (ABO21709), A/New Caledonia/20/1999 (ABF21272), A/California/04/2009 (ACP41105), A/WS/33 (ABD77796), A/WSN/33 (ABF47955), and A/NWS/33 (AAA92279). (B) Sequence alignment of H1 to H15 HAs. All HA sequences were obtained from the NCBI Influenza Virus Resource, and each HA subtype was aligned individually using Clustal W. For H5 and H7 HA alignments, sequences containing a polybasic cleavage site were not included. The consensus sequence of each subtype is shown. LPAI, low-pathogenic avian influenza virus.