Figure 5.

Generation of Hf09 NS_Sk02 virus reassortants. (a) Schematic diagram of genome composition of WT and mutant viruses. Hf09 NS_SK02 virus has the background of Hf09 WT (Blue) with NS segment replaced with that from Sk02 (Red) virus. Hf09 NS_Sk02 M2 mutant viruses (turquoise) are derivatives of Hf09 NS_SK02 wherein the 7th segment contains various mutations. (b) MDCK- M2 cells were infected with Hf09 NS_Sk02 WT and the respective Hf09 NS_Sk02 M2 mutant viruses at an MOI of 0.001. The supernatant collected at different time points was subjected to plaque assay. Viral protein expression from infected cells (MOI of 1, at 36 h.p.i.) was determined by Western blotting (lower panel) (p.f.u.—plaque forming unit; time p.i.—Time post infection).