Fig. 3.

Role of MRP2 5′UTR on translation efficiency of the luciferase reporter transcript in in vitro translation assays. a, schematic representation of the constructs used in in vitro translation assays. −247, −204, −204M1, −204M2, −99, and −99M1 are the same as described in Fig. 2a. C35, the fragment between T7 promoter and luciferase gene in T7 control DNA vector. b, in vitro translation assays of 5′UTR-luciferase transcript. The T7-5′UTR-luciferase cassette was amplified by PCR from the corresponding vectors as described in a; the cassettes were used as templates to synthesize the capped, 5′UTR-fused luciferase transcripts. The 5′UTR-luciferase transcripts (0, 2, 4, 10, 20, or 40 ng of mRNA) were added to rabbit reticulocyte lysate mixture. The lines are the best linear fit of the relationship between luciferase activity and mRNA transcript concentration. The slopes represent translation efficiencies. The statistical significance of the difference between the slopes was tested by Prism 4.0 (GraphPad Software, San Diego, CA).