miRNAs act through predicted MREs in the nAChR 3′ UTRs. Predicted MREs in the indicated nAChR 3′ UTRs were disrupted via site-directed mutagenesis (mutated nucleotides are shown in red) and assayed for miRNA-mediated repression of luciferase activity as described in Supplemental Figure S1. Each bar represents the normalized luciferase activity assayed from HEK293T cells transfected with the indicated luciferase/nAChR 3′-UTR construct and a miRNA precursor mimic, or negative control miRNA precursor mimic (n = 3–6). Data are presented as mean ± SEM. P-values compared with a negative miRNA control derived from the Student's t-test are indicated. The types of seed match sites (binding sites corresponding to the miRNA seed region) are indicated.
