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. 2013 Oct 1;8(10):e71568. doi: 10.1371/journal.pone.0071568

Figure 1. MiR-181a directly regulates TNF-α expression via 3’ UTR sites.

Figure 1

(A) Schematic of TNF-α mRNA and the luciferase reporter plasmids containing the miR-181a binding sites of TNF-α mRNA. The 3’ UTR sites were inserted downstream of the luciferase reporter, as indicated. TGAATGT was the predicted target site of miR-181a. (B) MiR-181a sequences and predicted binding site between miR-181a and TNF-α mRNA. Sequence of miR-181a (www.mirbase.org) is shown. TNF-α mRNA has one putative binding site for miR-181a on the 3’ UTR. Seven nucleotides of TNF-α 3’ UTR (underlined) were replaced with GACCGGT using site-directed mutagenesis in order to disrupt the binding with miR-181a seed regions. (C) CHO cells were transfected with each of the constructed plasmids, together with miR-181a and Renilla luciferase reporter plasmid (*P < 0.05, n = 8).