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. 2010 Oct 4;107(42):18161–18166. doi: 10.1073/pnas.1005595107

Fig. 4.

Fig. 4.

Essential roles of Mecp2-regulated miRNAs in regulation of Bdnf expression. (A) Schematic diagram of the pISO luciferase reporter containing the first 800 bp of the Bdnf 3′ UTR (pISO-Bdnf). Also shown are aberrantly up-regulated miRNAs (>1.5-fold) in KO cerebella at the early-symptomatic stage (6 wk postnatal) that are predicted to target the Bdnf 3′ UTR by at least two independent target-prediction algorithms (TargetScan, miRanda, or PicTar/DIANA microT). (B) Luciferase assays in 293T cells transfected with miRNA mimics duplexes (100 nM). Error bars represent SEM. *P < 0.05; n ≥ 4. Note that other miRNAs [cel-miR-67 (specifically expressed in C. elegans) and miR-137] were used as negative controls for pISO-Bdnf. (C) Luciferase assays in WT cerebellar neurons transfected with control (100 nM) or miR-381/495 (100 nM) 2’-O-Methyl oligonucleotide inhibitors. Error bars represent SEM. *P < 0.05; n ≥ 5. (D) Immunoblotting of Mecp2 in postnatal cortical neurons infected with lentiviruses expressing a control shRNA or a shRNA specific for Mecp2. (E) Bdnf ELISAs in postnatal cortical neurons. Error bars represent SEM. *P < 0.05; n = 4.