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. 2016 Jun 15;8(6):2727–2737.

Figure 2.

Figure 2

Validation that miR-503 targets the 3’-UTR of the Bmpr1a gene. A. Schematic of Bmpr1a mRNA and the luciferase reporter plasmids containing the miR-503 binding sites of Bmpr1a mRNA. The 3’-UTR sites were inserted downstream of the luciferase reporter and CGCTGCT was the predicted target site of miR-503. B. MiR-503 sequences and the predicted binding site between miR-503 and Bmpr1a mRNA. The sequence of miR-503 (www.mirbase.org) is shown. The Bmpr1a mRNA has one putative binding site for miR-503 on the 3’-UTR. The seed region is composed of seven nucleotides of the Bmpr1a 3’-UTR (red) and these were mutated using site-directed mutagenesis in Bmpr1a mRNA-muta. C. HEK293T cells were transfected with each of the constructed plasmids, together with the miR-503 luciferase reporter plasmid. D. HEK293T cells were transfected with a miR-503 mimic or inhibitor, together with the Bmpr1a 3’-UTR activated luciferase reporter plasmid (n=3; *p<0.05 vs. blank).