Fig. 2:

Fluorescence-based BRBV MG system in HEK293T/17 cells. A) Schematic of the segment 2-based BRBV MG plasmid encoding ZsGreen gene. PpolI: human RNA polymerase I promoter, TpolI: human RNA polymerase I terminator, NCR: non-coding region. BRBV MG activity is shown by B) fluorescence signals and C) fluorescence intensity with or without supplementation of PB2 helper plasmid at 72 hours post-transfection. D) Time course of BRBV MG activity at 24, 48, and 72 hours post-transfection. Fold induction of BRBV MG activity was obtained by normalizing PB2+ fluorescence intensity with PB2-. For B)-D), HEK293T/17 cells were seeded in 24-well plates and transfected with a MG-ZsGreen plasmid (0.25 μg) together with helper plasmids (0.1 μg each). E) BRBV MG activities induced by transfection of serially diluted helper plasmids at 72 hours post-transfection. HEK293T/17 cells were seeded in 96-well plates and transfected with a MG-ZsGreen plasmid (125 ng) together with helper plasmids (25 ng each as the initial amount which were further diluted for :2, :8, :16, :32, and :64). Data are representative of the average of three independent experiments or triplicate in one experiment. ***: p < 0.0005, ****: p < 0.00005.