HEK-293T cells were co-transfected with 200 ng psiCheck-2 target genes 3’ UTR luciferase reporter plasmid or psiCheck-2 mutant target genes 3’ UTR luciferase reporter plasmid, along with miRNA mimics or control (final concentration, 50 nM). After 24 h, cells were collected and assayed for both firefly and Renilla luciferase activities using the Dual-Luciferase Reporter System following the manufacturer’s instructions (Promega, Madison, WI, USA). The results were calculated as the ratio of Renilla luciferase activity to firefly luciferase activity and recorded as the mean + SD from three replicate wells.
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