FPV-T7-driven intracellular transcription and translation from transfected linear DNA templates of NSP2 and NSP5.

<p>COS-7 cells infected with FPV-T7 Pol for 1 hour and transfected with 1 µg of linear PCR amplicon encoding either RV segments 8 or 11 or EGFP. pcDNA3-NSP2 and pT7V-NSP5 were used as protein controls. Cells were incubated for 24 hours, prior to fixing and staining for RV proteins as previously described. Cells transfected with EGFP were only stained with Hoechst 33342. Samples were imaged using a fluorescence microscope. All panels were exposed to UV light, panels A, B, C, and D were exposed to blue and panels E and F were exposed to green wavelengths of light. Panel A: mock transfection; panel B: transfection with T7EGFP; panel C: transfection with pcDNA3-NSP2; panel D: transfection with T7RFS8; panel E: transfection with pT7V-NSP5; panel F: transfection with T7RFS11. Cell nuclei were stained with Hoechst 33342. Scale bars: 100 µm.