MiR-128 and miR-137 synergize to reduce NF1 reporter protein expression.

<p>HEK293 cells were co-transfected with both the reporter gene (0.2 µg/reaction) and variable concentrations of pri-mir-128 and pri-mir-137 expression vectors. Luciferase activity was measured 48 hours later. (<b>A</b>) pri-mir-128 plasmid concentration was kept at 0.4 µg/reaction while pri-mir-137 concentration varied from 0.1–0.4 µg/reaction. (<b>B</b>) pri-mir-137 plasmid concentration was kept at 0.4 µg/reaction while pri-mir-128 concentration varied from 0.1–0.4 µg/reaction. Scramble 2 plasmid was supplemented in some of the reactions to achieve final miRNA plasmid concentration of 0.8 µg/reaction. These assays demonstrated that miR-128 and miR-137 synergistically reduce NF1 reporter expression when levels of the less abundant miRNA in the reaction are 50% or higher of the more abundant miRNA. The average value of two single miRNA expression plasmids (combination of scramble 2 plus pri-mir-9 or pri-mir-181 or pri-mir-218) predicted not to bind <i>Nf1</i> 3′-UTR were used as controls. Data show the mean ± s.e.m from 4 independent transfections (*, P<0.05; **, P<0.01, ***, P<0.001).