miR-181a Post-Transcriptionally Downregulates Oncogenic RalA and Contributes to Growth Inhibition and Apoptosis in Chronic Myelogenous Leukemia (CML)
Figure 2
miR-181a-directed repression of Renilla luciferase reporter genes containing RalA-3′-UTR segments.
K562 cells were cotransfected with scrambled RNA or miR-181a together with RalA-3′-UTR or RalA-mut-3′-UTR luciferase reporter in the presence of firefly luciferase reporter plasmid as indicated. Renilla luciferase activity and firefly luciferase activity were measured by dual-luciferase reporter assay (Promega). Renilla luciferase activity was normalized to firefly luciferase activity. The data represent the mean value of three independent experiments. (A) overexpression of miR-181a significantly repressed Renilla luciferase activity in cells transfected with RalA-3′-UTR, but not RalA-mut-3′-UTR vector; *p<0.01. (B) Human RalA-3′-UTR and its miR-181a target site predicted by TargetScan software. Mut: contains 7-base-mutation at the miR-181a target region.