FIGURE 7.

The effects of poly(A) tail length on IRES-mediated translation efficiency. (A) In vitro transcribed RNAs (0.8 μg) derived from linearized monocistronic reporter plasmids pDHAV-1-An containing various lengths of the poly(A) tail (n: 0, 5, 10, 15, 20, 25, 30, and 40) were co-transfected into DEFs with 0.1 μg of pGMLR-TK plasmid. Cell lysates were collected at 4, 6, 12, and 24 hpt to measure FLuc/RLuc activity. Columns and bars represent means and standard deviations, respectively, of three independent transfections. ^∗∗∗P < 0.001, ^∗∗P < 0.01. (B) The RNA-launched infectious clone pR-DHAV-1-An was transfected into DEFs, and the cell lysates were collected at 4 hpt for western blot analysis with anti-DHAV-1 monoclonal antibody 4F8 and HRP-labeled goat anti-mouse antibody. The grayscale value in poly(A)₂₅ group was set as 100%.