Figure 5. miRNA-sensor assay using luciferase expression as an indicator of miRNA activity after transfection of HeLa cells with various constructs. (A) Relative luciferase expression after transfection of HeLa cells with triplet-cassette constructs: pGL4-10–an empty pGL4-10 vector for standardization control, pGL4-10 + wt-Triplet–vector containing a wild-type triplet cassette containing potential binding sites for c-miR-Ch9, pGL4-10 + mut-Triplet–a vector containing a triplet cassette with mutated binding sites for c-miR-Ch9. (B) Relative luciferase expression after transfection of HeLa cells with 3′UTR constructs. pGL4-10–an empty pGL4-10 vector for standardization control, pGL4-10 + wt-3′UTR–vector containing a wild-type 3′UTR containing a single potential binding site for c-miR-Ch9, pGL4-10 + mut-3′UTR–a vector containing a single mutated potential binding site for c-miR-Ch9. (C) The pGL4-10 + wt-Triplet cassette transfection was repeated with concurrent addition of anti-LNA for our c-miR-Ch9. (D) An average over all transfection experiments performed (total of 5 experiments with 3 triplicates for every condition).