Table 3 Mutations observed in antibody-resistant isolates.

Gene [segment] (reference sequence, accession)	CalX wt Change: AA [NT]	P2 Change: AA [NT]	P16A1 Change: AA [NT]
PB2 [segment 1] (A/Puerto Rico/8/34, V00603.1)¹³	G309D [G953A], Y360S [A1106C], [C1540A], [G1815A]	G309D [G953A], Y360S [A1106C], [C1540A], [G1815A], A661V [C2009T]¹⁴	G309D [G953A], Y360S [A1106C], [C1540A], [G1815A], A661V [C2009T], E676D [A2058C]¹⁵
PB1 [segment 2] (A/Puerto Rico/8/34, J02151.1)	A53G [C282G], [T504A], S216G [A670G], *[C730T]*¹⁶, L300F [G924C], H473L [A1442T], S517I [T1563G], [T1574G]	A53G [C282G], [T504A], S216G [A670G], L300F [G924C], H473L [A1442T], S517I [T1563G], [T1574G]	A53G [C282G], [T504A], S216G [A670G], L300F [G924C], H473L [A1442T], S517I [T1563G], [T1574G]
PA [segment 3] (A/Puerto Rico/8/34, V01106.1)¹³	K158R [A497G], *K391E [A1194G]*, [T1308A]	K158R [A497G], [T1308A]	K158R [A497G], [A783G], [T1308A]
HA [segment 4] (A/California/07/2004, EU103820.1)	100% homology	100% homology	100% homology
NP [segment 5] (A/Puerto Rico/8/34, J02147.1)¹³	[G306A], D247N [G784A]	[G306A], D247N [G784A]	Failed to amplify¹⁷
NA [segment 6] (A/California/07/2004, EU103978.1)	100% homology	100% homology	V240I [G718A]
M1 &M2 [segment 7] (A/Puerto Rico/8/34, V01099.1)	100% homology	100% homology	100% homology
NS1 & NS2/NEP [segment 8] (A/Puerto Rico/8/34, J02150.1)	100% homology	100% homology	100% homology

¹³ CalX wt and antibody-selected isolates most closely related to A/reassortant/NIBRG-14(Viet Nam/1194/2004 × Puerto Rico/8/1934)(H5N1).
¹⁴ Mutations present in P2 and/or P16 but not in wild type are shown in bold type
¹⁵ Present as a mixture with wild type sequence at a mutant:wildtype ratio of approximately 1:2 (see Figure S3).
¹⁶ Mutations present only in wild type are shown in bold italic type.
¹⁷ See Figure S3.

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