Figure 7.

Luciferase reporter for Ank3 3′-UTR shows regulation by miRs. (A) Sequential luciferase signal was recorded from a dual luciferase reporter incorporating Ank3 3′-UTR transfected into mCCD cells, seeded onto filter supports, and normalized to empty vector controls (100%). The Ank3 3′-UTR signal alone cotransfected with control LNA was reduced compared with the empty vector. Expression of each miR was reduced by cotransfection of 50 nM LNA (as in Figure 4), and luciferase reporter expression increased significantly with reduction of miR-335–3p expression (n=9). (B) In the converse experiment to A, miR expression was increased using miR mimics and pre-miR plasmids (as in Figure 5), and a further, significant (P<0.05) reduction in normalized luciferase expression was observed in cells overexpressing miR-335–3p (n=9).