After culturing for 24 hours, 293T cells were co-transfected with BANCR 3′-untranslated
region and cloned into the psiCHECK-2 vector and miR-mimics or NC mimics (100 ng/well)
using Lipofectamine 3000 (Invitrogen). After 48 hours, the cells were washed using PBS and
then lysed with passive lysis buffer at room temperature for 20 minutes. The incubation
lysates were collected and transferred to 96-well plates, and the aliquots were added to
the plates. The firefly luciferase activity was measured in the Infinite M200 plate reader
(Tecan, Männedorf, Switzerland) system after adding Luciferase Assay Reagent II
immediately. Next, the Stop & Glo reagent was used to initiate the Renilla luciferase.
The relative luciferase activity for miRNA-195-5p is shown relative to the NC mimics.
region and cloned into the psiCHECK-2 vector and miR-mimics or NC mimics (100 ng/well)
using Lipofectamine 3000 (Invitrogen). After 48 hours, the cells were washed using PBS and
then lysed with passive lysis buffer at room temperature for 20 minutes. The incubation
lysates were collected and transferred to 96-well plates, and the aliquots were added to
the plates. The firefly luciferase activity was measured in the Infinite M200 plate reader
(Tecan, Männedorf, Switzerland) system after adding Luciferase Assay Reagent II
immediately. Next, the Stop & Glo reagent was used to initiate the Renilla luciferase.
The relative luciferase activity for miRNA-195-5p is shown relative to the NC mimics.