NS1 p85β-binding variants have arisen in human IAVs. (A) Sequence alignment of NS1 proteins from early human H1N1 IAVs highlighting the emergence of 95I. BM18, A/Brevig Mission/1/1918 (H1N1); Melbourne/35, A/Melbourne/1935 (H1N1); Henry/36, A/Henry/1936 (H1N1); Hickox/40, A/Hickox/1940 (H1N1); Bel/42, A/Bel/1942 (H1N1); Weiss/43, A/Weiss/1943 (H1N1). (B) Graph indicating the evolution of residues 95 and 145 in H1N1 IAVs (excluding the 2009 pandemic virus). H1/Brisbane/07, A/Brisbane/59/2007 (H1N1). The percentage of isolates harboring a particular residue within the indicated period is plotted. n, the number of isolates analyzed per time point; *, results for the years 2006 to 2008 (data are for 708 isolates). Selected strains are highlighted. (C and D) Same as panel B, but for H3N2 (C) and 2009 pandemic H1N1 (D) viruses. H3/Boston/13, A/Boston/DOA2-172/2013 (H3N2); H3/Brisbane/07, A/Brisbane/10/2007 (H3N2). **, results for the years 2014 and later (data are for 2,200 isolates).