Influenza A viruses with avian NA promptly adapt to cleave DAF.

A: As showed in Fig 10I, HEK293T cells were transfected with eight different avian derived NAs. B: To produce reverse genetics (RG) reassortant A/Puerto Rico/8/34 (PR8) viruses, HEK293T cells were transfected with seven plasmids encoding segments 1–5, 7 and 8 from PR8, and the segment 6, which encodes NA, from the indicated viruses. C: After one round of amplification in embryonated chicken eggs, recovered viruses were titrated. D-E: A549 cells were infected with PR8 NA-H5N2 at multiplicity of infection (MOI) of 3 and samples collected at the indicated timepoints to titrate released virions (D) and analyze DAF cleavage by western blot (E) (D: pooled data from three independent experiments; E: representative blot from three independent experiments). Statistical analysis detailed in materials and methods.

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