Fig. 5

miR-132 suppresses Nurr1 expression post-transcriptionally. a The putative site in the Nurr1 3′-UTR that contains the ‘seed sequence’ for miR-132 binding. The g and a shown in red indicate the miR-132 binding site in the mutant form of Nurr1. mGCs were transfected with 100 nM miR-132 mimics, miR-132 inhibitors or the corresponding negative controls. Cells were co-transfected with the wild-type (WT) Nurr1 3′-UTR luciferase reporter plasmid (b), the mutant (MU) Nurr1 3′-UTR luciferase reporter plasmid (c) or the NBRE-luciferase reporter plasmid (d). After 48 h, luciferase assays were performed, and the results were normalized using constitutive Renilla luciferase. e Western blot analysis of Nurr1 and Nur77 protein expression in mGCs 48 h after transfection with 100 nM of miR-132 mimics, miR-132 inhibitors or the corresponding negative controls. The upper panels depict representative Western blots, and the lower panels present the statistical summary of the densitometric analysis from three independent experiments, indicating expression levels relative to negative controls after normalization to GAPDH. f Real-time PCR analysis of Nurr1 mRNA levels in mGCs 48 h after transfection. The results represent the mean +/-SEM of three independent experiments performed in triplicate. **p < 0.01; # p < 0.05, compared with the negative control (NC)