MiR-330 negatively regulated SH3GL2 through binding to the 3′-UTR of SH3GL2.

<p>(A) Schematic diagram of putative miR-330 binding site in the 3′-UTR of SH3GL2 in human. The seed sequence of miR-330 matches 3′-UTR of SH3GL2 (in bold). (B) The corresponding mutated nucleotides of the SH3GL2 3′-UTR was labeled in bold bellow. (C) Relative luciferase activities of SH3GL2 wild type 3′- UTR were obtained by co-transfection of PBS (mock), negative control miRNA or miR-330 precursors; and calculated as the ratio of firefly/renilla activities in the cells and normalized to those of the control. The results were presented as mean±SD from three independent experiments with each experiment in triplicate. Asterisk indicates significant difference when compared to control (P<0.05). (D) Relative luciferase activities of pmirGLO plasmid, SH3GL2 wild type and mutant type 3′- UTR were obtained by co-transfection of miR-330 precursor; and calculated as the ratio of firefly/renilla activities in the cells and normalized to those of the control. The results were presented as mean±SD from three independent experiments with each experiment in triplicate. Asterisk indicates significant difference when compared to control (P<0.05).