Research Paper Volume 12, Issue 15 pp 15414—15435

Figure 7. Validation of direct targeting between the partners within the circLDLR-miR-1294-CYP19A1 ceRNA network. (A) The sketch map of LDLR gene and its derived circLDLR (has_circ_0006877). The circLDLR is formed by backsplicing junction of the 13 and 14 exons of LDLR. (B) The RNase R tolerance test proved the circular format of circLDLR. ACTB was used as the negative control. (C) Three binding sites of miR-1294 on circLDLR predicted by miRanda software. (D) The binding site of miR-1294 on CYP19A1 predicted by miRanda software. (E) The relative luciferase activity was measured following co-transfection of miR-1294 mimics with the constructs encoding the wild-type or mutant circLDLR cDNA into the KGN cells. (F) The relative luciferase activity was measured following co-transfection of miR-1294 mimics with the constructs encoding the wild-type or mutant CYP19A1 3’-UTR into the KGN cells. The firefly luciferase activities were normalized by renilla luciferase activities. The experiments were performed independently in triplicates. * indicates p < 0.05, ** indicates p < 0.01, NS indicates no significant difference. The results are presented as means ±SEM.