Fig. 1

miR-9-5p regulated expression of both REST and MOR by targeting REST in primary mouse bone marrow mesenchymal stem cells (BMSCs). a DNA sequence of a predicted binding site of mmu-miR-9-5p within the wild-type 3′-UTR region of REST mRNA as well as a mutated 3′-UTR of REST was cloned into a luciferase reporter construct, respectively. The corresponding mRNA sequence was shown. b miR-9-5p suppressed the luciferase activities expressed by WT-REST but not MUT-REST luciferase construct. Luciferase activities were measured at 48 h post transfection. c Expression efficacy of miR-9-5p mimics was verified using qRT-PCR at 24-h post transfection. d, e Overexpression of miR-9-5p decreased REST but increased MOR mRNA levels 24 h post transfection. f Accordingly, overexpression of miR-9-5p decreased REST but increased MOR protein expression 24 h post transfection. g Quantification of protein expression presented in f. qRT-PCR was used to evaluate mRNA level, and western blotting was used to evaluate protein expression. Mean ± SD is used to present all data. *p < 0.05, **p < 0.01, and ***p < 0.001