Figure 4

Position of mismatches in the inhibitor target site affects inhibitor functionality. The effect of mismatches on inhibitor function was determined for (a) miR-21 in HeLa cells, (b) for miR-122 in Huh-7 cells and (c) for miR-22 in HeLa cells; cell lines were chosen for high expression of the respective miRNAs. Transfections were performed 1 day after plating into 96-well plates, 10,000 cells/well, in antibiotic-free media. For each microRNA, the appropriate dual-luciferase reporter, at 100 ng/well, was co-transfected with either a fully matched inhibitor or one of a set of 11 mismatched inhibitors at concentrations from 0.03 to 20 nM. Dual-luciferase ratios were measured 2 days post-transfection. Results shown are averages from triplicate wells, normalized to appropriate controls, then expressed as fold-inhibition relative to transfection with a negative control. Error bars are ± 1SD (sample) of the original triplicate data, scaled for all subsequent calculations.