Fig. 3.

Simultaneous binding of miR-760, miR-186, miR-337-3p and miR-216b to CK2α 3′-UTR is necessary for suppression of CK2α expression in IMR-90 cells. (A) The luciferase reporter vector (pGL3) containing wild-type (WT) or mutant 3′-UTR of the CK2α. Individual miR-binding sites were mutated in the CK2α 3′-UTR vector. (B) Mutant vectors (m1, m2, m3, and m4) were generated using designed mutagenic oligonucleotide primers. Mutations in the core sequences are underlined. Watson-Crick and wobble base (G-U) pairings are indicated by solid and dashed vertical lines, respectively. (C) Luciferase reporter vectors, either WT or mutant (m1, m2, m3, and m4) were co-transfected with either control miRNAs or all four miRNA mimics into IMR-90 cells (PDL 33) at a final concentration of 100 nM. Luciferase activity was measured 24 h after transfection and normalized to Renilla. Data are shown as the means ± SEM. *P < 0.05.