Pmirglo dual luciferase mirna target expression reporter

Bioinformatics was utilized to predict the potential binding sites between 3′-UTR of OTUD7B and miR-103. The OTUD7B 3′-UTR wild sequence or OTUD7B 3′-UTR mutant sequence was inserted into the pmiRGLO dualluciferase miRNA target expression reporter (Promega, Madison, WI, USA). 293T cells were seeded on 24-well plates (5 × 10 5 cells / well) and 24 h later, the cells were co-transfected with luciferase reporters and miR-103 mimic or negative control mimic using with Lipofectamine® 2000 (Invitrogen, Carlsbad, CA, USA). The luciferase activity was detected 48 h after transfection with Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA) in accordance with the manufacturer's instructions. NF-κB luciferase reporter plasmid (Beyotime Institute of Biotechnology, Nantong, Jiangsu, China) with OTUD7B plasmid or OTUD7B shRNA were co-transfected into NHL using Lipofectamine® 2000 (Invitrogen, Carlsbad, CA, USA) according to manufacturer's instructions. After 24 h, luciferase activity was measured using the luciferase kit (Beyotime, Hangzhou, China).