Figure 2.

JAM-A is a target gene of miR-495 in breast cancer cells. (A) Schematic illustration of the conserved miR-495 binding sites. The JAM-A 3′-UTR contains one predicted miR-495 binding sites. The seed regions of miR-495 and the seed-recognizing sites in the JAM-A 3′-UTR are indicated in red, and all nucleotides in seed-recognizing sites are completely conserved across several species. (B) Western blotting analysis of JAM-A protein levels in MCF-7 and MDA-MB-231 cells transfected with miR-495 mimic or inhibitor. (C) Direct recognition of the JAM-A 3′-UTR by miR-495. Firefly luciferase reporters containing either wild-type (WT) or mutant (MUT) miR-495 binding sites in the JAM-A 3′-UTR were co-transfected into MDA-MB-231 cells with the scrambled negative control RNA, miR-495 mimic or inhibitor. At 24 h post-transfection, the cells were assayed using a luciferase assay kit. The results were calculated as the ratio of firefly luciferase activity in the miR-495-transfected cells normalized to the control cells. *P < 0.05; **P < 0.01