TargetScan was used to predict the potential targets of miR-421 [18 ]. The 3’-UTR of BMF containing miR-421 binding sites was cloned into pMIR vectors (Ambion, USA) to generate the BMF wild-type (BMF-WT) or BMF mutated (BMF-MUT) plasmid. For the reporter activity analysis, 293 T cells were cotransfected with BMF-WT or BMF-MUT plasmids and miR-421 mimic or mimic control using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer’s protocol. After 24 h, luciferase activity was measured using the Dual-Luciferase Reporter Assay System (Promega, USA) [19 (link)].
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