Figure 5

Mismatched inhibitors show similar positional effects on the expression of an endogenous target. The effect of mismatches on miR-122 inhibitor function was determined by measuring steady-state levels of endogenous ALDOA mRNA. The set of miR-122 inhibitors [11 mismatched and one fully matched, and a negative control (NC) inhibitor] were transfected into Huh-7 cells 1 day after plating into 96-well plates, 10,000 cells/well, in antibiotic-free media. Media was changed to new, antibiotic-free media approximately every 3 days. Response of ALDOA mRNA levels relative to a housekeeping gene was measured on day 6 (white bars) and day 9 (grey bars) post-transfection by branched-DNA assay. Results shown are averages from triplicate wells transfected with 4 nM inhibitor, normalized to appropriate controls, then expressed as fold-inhibition relative to transfection with the negative control. Error bars are ± 1SD (sample) of the original triplicate data, scaled for all subsequent calculations.