Research Paper Volume 12, Issue 4 pp 3218—3237

Figure 5. LINC00623 competes with HRAS for miR-101 binding. (A) Four different types of luciferase reporter gene vectors were constructed: a wt-LINC00623 or a wt-HRAS 3′-UTR containing the wild-type miR-101 binding site, and a mut-LINC00623 or a mut-HRAS 3′-UTR containing the mutated miR-101 binding site. (B, C) These vectors were cotransfected into HEK293 cells with miR-101 mimics or miR-101 inhibitor; the luciferase activity was examined. (D, E) RIP assays were performed to confirm the predicted binding between LINC00623 and miR-101 using an anti-AGO2 antibody. (F) the HRAS luciferase reporter vector, miR-101 mimics and wt- or mut-LINC00623 vector were cotransfected into HEK293 cells, the luciferase activity was examined. The data are presented as mean ± SD of three independent experiments. *P<0.05, **P<0.01.