Fig. 1.

The 3′-UTR of Pax3 is a target of miR-27. (A) The predicted interaction includes a perfect 8-base pair match to the seed region of the miRNA (dark gray). Additional features that would boost efficacy of the target interaction include a flanking AU-rich sequence (underlined). (B) Transfection of 293 cells, expressing the psicheck-2 luciferase vector containing the Pax3 3′-UTR downstream of the Renilla luciferase (R. luc) sequence, with miR-27b precursors, resulted in reduced R. luc activity that was lost on mutation of the miR-27 target site (Pax3–3′UTRmut). Conversely, transfection with a miR-27b inhibitor (anti-miR-27b), which blocks the activity of endogenous miR-27b in 293 cells (Fig. S2), resulted in increased R. luc activity. Reported results are normalized to firefly luciferase activity, which serves as an internal transfection control, and presented relative to R. luc activity after transfection with random, nontargeting, control oligonucleotides. Results are from 3 independent experiments.