Fig. 2

In vivo targeting of Hcrt by miR-137-3p and miR-665-3p. (A) Schematic representation of ICV injection in P4 mouse pups. (B) Injection of a mixture of mmu-miR-137-3p/miR-665-3p mimics (n = 9) and anti-miR-137-3p/anti-miR-665-3p (n = 7) in P4 pups. Littermate controls were injected with a scramble (n = 16 per group) or an antiscramble (n = 15). RNA was isolated and analyzed by qRT-PCR. Data are presented as the magnitude of the difference compared with controls ± SEM. HCRT-1 peptide was extracted from the brains and analyzed by radioimmunoassay (RIA) (upregulation by miRNA mimic, n = 8 vs. controls n = 7; downregulation by anti-miR, n = 15 vs. controls n = 7). Counts are normalized to control and presented as mean ± SEM. Pcbd: Pterin-4-alpha-carbinolamine dehydratase function as the control since it is not a target of mmu-miR-137-3p/miR-665-3p. miR-155 is included as a control to illustrate that miRNAs in general are not affected by anti-miR injections. *P < 0.05; **P < 0 0.01; ***P < 0.001, two-tailed Student’s t test.