Luciferase Reporter Assay for miR-580 Targets

The wild-type (WT) or mutant-type (MT) fragments of SATB1-AS1 or OAS2 3ʹuntranslated region (3ʹUTR) containing miR-580 target sequences were synthesized and inserted into the pGL3 promoter vector (Promega). These sequences were named SATB1-AS1-wt, SATB1-AS1-mut, OAS2 3ʹ-UTR-wt and OAS2 3ʹ-UTR-mut, respectively. The above vectors, miR-580 mimic or mimic control were co-transfected into 293T cells by using Lipofectamine 2000 (Invitrogen). The cells were collected 48 h after induction. The luminescence was assessed with a dual luciferase reporter gene detection system (Promega). The results were then normalized to Renilla luciferase activity [25 (link)].