Fig. 2

Structure and functional characterization of the putative miR-17-5p/miR-106b-5p target identified in the TRIM8 3’UTR sequence. a Schematic representation of the pMIR luciferase reporter construct containing the TRIM8 3’UTR sequence (wild-type or mutated) cloned downstream the Luciferase gene. Below it is shown the sequence alignment between the miR-17-5p/miR-106b-5p “seed” sequence and the TRIM8 3’UTR, as well as the evolutionary conservation across species. b, c, d, e Luciferase assays. The HK-2 and HCT116 cells were transfected with Negative Control miRNA Mimic, miR-17-5p or miR-106b-5p (alone or together), anti-miR-17-5p or anti-miR-106b-5p (alone or together), along with pMIR luciferase reporter construct containing TRIM8 3’UTR (wt or mut). Cells were lysed and luciferase activity was determined as described in the Material and Methods section. Transfection efficacy was normalized by Renilla Luciferase activity. Data represent the averages of at least three independent experiments with their standard deviations. ** p-value < 0.005; *** p-value < 0.001