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. 2014 Oct;88(20):12006–12016. doi: 10.1128/JVI.01847-14

FIG 2.

FIG 2

Viral rescue of X31-sciIV by reverse genetics. A mixture of 293T cells and X31 HA-expressing MDCK cells was cotransfected with pPolI X31 NA, pPolI HA(45)GFP(80), pCAGGS X31 HA, and ambisense pDZ rescue plasmids encoding PR8 PB2, PB1, PA, NP, M, and NS. (A) At 48 h posttransfection, tissue culture supernatants were harvested for infecting X31 HA-MDCK cells and, at 48 h postinfection, GFP expression was observed using fluorescence microscopy. Images are shown at ×40 magnification. Scale bar, 10 μm. (B) Rescued X31-sciIV virus was then plaque purified, and the viral growth on parental and X31 HA-expressing MDCK cells (MOI = 0.001) was compared to that of WT X31 virus using a hemagglutination assay.