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. 2016 Aug 2;9(39):25414–25426. doi: 10.18632/oncotarget.11003

Figure 3. APE1 expression is inhibited by miR-513a-5p.

Figure 3

(A) The graph depicts the construction strategy of GLO-APE1-513 (WT) and GLO-APE1-513M (MT). The APE1-3′UTR sequence (258 bp) containing the putative miR-513a-5p binding site and mutant sites was synthesized and inserted respectively into the pmirGLO Dual-Luciferase miRNA target expression vector at XhoI and Xbal restriction enzyme sites. (B) The effects of miR-513a-5p binding of the 3′UTR of APE1 on the luciferase activity. HOS cells were transfected with GLO-APE1-513 vector and GLO-APE1-513M vector. Luciferase activity was detected at 48∼72 h after transfection with or without scrambled mimics, miR-513a-5p mimics and miR-513a-5p inhibitor, respectively. An asterisk (*) indicates that the difference between the marked group and the scramble-transfected group is statistically significant (p < 0.01). (C) APE1 protein expression was detected by Western blot after transfection with scrambled mimics, miR-513a-5p mimics and miR-513a-5p inhibitor in HOS cells. β-actin was used as the internal control. Both representative blots and statistical analyses (D) are shown. An asterisk (*) indicates that the difference between the marked group and the scramble-transfected group is statistically significant (p < 0.01).