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. Author manuscript; available in PMC: 2018 Jul 31.
Published in final edited form as: RNA Biol. 2009 Jan 5;6(1):84–89. doi: 10.4161/rna.6.1.7572

Figure 2.

Figure 2

The 472 nucleotide p27Kip1 5′-UTR contains the highest IRES activity. Direct RNA transfections of bicistronic dual luciferase mRNAs containing the p27Kip1 5′-UTRs were performed using DMRIE-C transfection reagent. Cells were harvested ten hours after transfection followed by assays for Renilla and firefly luciferase activity levels (in relative light units) and calculation of the firefly/Renilla ratio. (A) Transfection of MDA-MB-231 cells with RNAs containing the 677, 472 and 403 nucleotide p27Kip1 5′-UTRs. A negative control transfection with an RNA transcript containing the reverse complement of the 472 nucleotide 5′-UTR was also performed (472R). Asterisks indicate statistically significant reduction in firefly/Renilla ratio relative to the 472 nucleotide 5′-UTR (unpaired t test; p = 0.0022 for the 403 nucleotide 5′-UTR; p = 0.00021 for the 677 nucleotide 5′-UTR). (B) Transfection of MCF7 cells with RNAs containing the 472 and 575 nucleotide p27Kip1 5′-UTRs and the 472R negative control. Asterisk indicates statistically significant reduction in firefly/Renilla ratio relative to the 472 nucleotide 5′-UTR (unpaired t test; p = 0.03 for the 575 nucleotide 5′-UTR).