Investigating MEG3-miR-29c Regulatory Axis

Wide type (WT)-MEG3 containing binding sites were amplified by PCR using Ultra HiFidelity PCR Kit (TIANGEN, Beijing, China), and mutation type (MUT)-MEG3 was synthesized by Sangon (Shanghai, China). The obtained MUT-MEG3 and WT-MEG3 sequences were cloned into pmirGLO vectors. The vectors carried with WT-MEG3 were co-transfected with miR-29 c mimics, miR-29 c inhibitors, or miR-NC into MRC-5 cells via lipofectamine 3000 ((Life Technologies, Carlsbad, USA). The co-transfection procedures of MUT-MEG3 were consistent with that of the WT-MEG3 group. Luciferase activities were surveyed 48 hours after transfection utilizing the Dual-Glo luciferase system kit (Promega, Madison, USA) [20 (link)].