Figure 3

Comparison of the activities of the sense and antisense IBDV MG. A and B Schematic representations of IBDV antisense hRB-Rluc (A) and hRB-GFP (B) reporter plasmids for the MG based on the B segment backbone. hPol I (red) and T (blue) represent the human Pol I promoter and mouse terminator I. C HEK293T cells (6-well plate format, 10⁶ cells/well, triplicate) were transfected with Fluc (0.01 µg/well), hB-Rluc or hRB-Rluc (2 µg/well), together with pVP1 and pVP3 (1 µg/well each). At 72 h post-transfection, the cells were lysed in passive buffer, and the Rluc and Fluc activities were measured using a multiplate reader. The expression of VP1, VP3, and GAPDH was analysed by western blotting. The data are presented as the means ± SD and are representative of three independent experiments. **, p < 0.01. D and E HEK293T cells (6-well plate format, 10⁶ cells/well, triplicate) were cotransfected with hB-GFP or hRB-GFP plasmids alone or together with pVP1 and pVP3 plasmids. Fluorescence microscopy (D) and western blotting (E) were performed to determine the expression of EGFP and viral proteins using specific antibodies. GAPDH was used as a loading control. Scale bar, 200 μm. Magnification, ×10.