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. 2010 Oct 20;85(1):357–365. doi: 10.1128/JVI.01694-10

FIG. 5.

FIG. 5.

Viral RNA polymerase activity of PB2-WT, PB2-E158G, and PB2-E627K in human cells. HEK-293T cells were transfected with a pPolI-NS-Luc plasmid (pBZ81A36) that expresses negative-sense virus-like RNA encoding a destabilized firefly luciferase enzyme that can be transcribed by the viral RDRP. The HEK-293T cells were also cotransfected with plasmids expressing NY1682 PB1, PA, and NP and one of the PB2 clones (WT, E158G, or E627K) to generate different viral RDRPs. Cells were also cotransfected with a Renilla luciferase expression plasmid to control for transfection efficiency. Eighteen hours posttransfection, both firefly and Renilla luciferase production levels were measured, and Renilla expression was used to normalize the data. The averages of triplicate experiments are shown, with error bars that represent SD. *, PB2-E158G had significantly greater activity than either PB2-WT (P = 1.6 × 10−5) or PB2-E627K (P = 4.2 × 10−5) when analyzed using a t test with unequal variance and Bonferroni's correction.