FIG 3.

miR-155 interacts with the NPPA 3′ UTR in an rs61764044 allele-specific manner. (A) miR-155 reduced the activity of the WT-Luc construct but not the 155 Mut-Luc construct (containing a 6-nucleotide mutation at the predicted miR-155 seed binding site) or the rs61764044 Minor-Luc construct. The ratio of firefly luciferase activity to renilla luciferase activity was normalized to the ratio in COS-7 cells transfected with constructs encoding luciferase without the NPPA 3′ UTR and renilla. For each construct (WT-Luc, 155 Mut-Luc, or rs61764044 Minor-Luc), the effect of the miRNA was compared to that of a scrambled negative-control miRNA mimic (relative luciferase activity). (B) Anti-miR-155 increased the activity of the WT-Luc construct but not the 155 Mut-Luc construct or the rs61764044 Minor-Luc construct. The ratio of firefly luciferase activity to renilla luciferase activity was normalized to the ratio in COS-7 cells transfected with constructs encoding luciferase without the NPPA 3′ UTR and renilla. For each construct (WT-Luc, 155 Mut-Luc, or rs61764044 Minor-Luc), the effect of the anti-miR was compared to that of a scrambled negative-control anti-miRNA (relative luciferase activity). The rs61764044 Minor-Luc construct contains the minor alleles of both rs61764044 and rs5068. Data are means ± SEMs (n = 6 biological replicates; two-tailed independent t test).