Fig. 3.

The size of the 3′UTR affects the ability of miR-27a to attenuate Runx1 expression. HEK293 cells were co-transfected with pRL-S3′UTR or pFF-L*3′UTR and increasing amounts of miR-27a or miR-NC (transfection control). Numbers at the x axis [arbitrary units (AU) 1–5] correspond to increasing concentrations of miR-27a as follows: 0 nM, 0.1 nM, 0.3 nM, 1 nM, and 3 nM. The effect of miR-27a on reporter activity was calculated relative to the corresponding miR-NC transfected cells (defined as 1.0). Reporter expression was normalized to internal control luciferase activity. The slope values (−0.096 and −0.157 for L*3′UTR and S3′UTR, respectively) indicate that the S3′UTR-dependent translation is more sensitive than L*3′UTR-dependent translation to inhibition by miR-27a. Of note, a square r of ≈0.93 indicates the close relationships between the values comprising the slopes. Data shown are representative of at least two independent experiments done in triplicate. Error bars indicate SD.