Oligoribonucleotides for the regulation of miRNA expression were obtained from GenePharma (Shanghai, China). mmu-miR-21 mimics (miR-21) were simple-stranded RNA oligonucleotides with similar biological functions to mature endogenous miR-21, and mmu-miR-21 inhibitors (anti-miR-21) were designed to down-regulate endogenous miR-21 expression. The sequences for each oligonucleotide are listed in Table I . Both oligonucleotides had their own negative control (NC). Entranster™ R4000 (Engreen Biosystems, Beijing, China) was utilized according to the manufacturer's instructions. The transfection assay was conducted based on a previously described protocol (26 (link)).
The C2C12 cells and MEFs were transfected with 100 nM of either a non-targeting small RNA oligonucleotide as a negative control (miR.NC) or miR-21 mimic, or mmu-miR-21 inhibitor in a well in a 24-well plate or 500 nM of the above in a well in a 6-well plate. The transfection efficiency was determined under a fluorescence microscope.
The C2C12 cells and MEFs were transfected with 100 nM of either a non-targeting small RNA oligonucleotide as a negative control (miR.NC) or miR-21 mimic, or mmu-miR-21 inhibitor in a well in a 24-well plate or 500 nM of the above in a well in a 6-well plate. The transfection efficiency was determined under a fluorescence microscope.