Scramble control sirna

Human hepatocellular carcinoma cell lines (HepG2) were purchased from the Institute of Biochemistry and Cell Biology of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, NY, USA), supplemented with 10% fetal bovine serum (FBS, Gibco) as well as 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen, CA, USA) and maintained at 37 °C with 5% CO2.
Small double-strand interference RNAs (siRNAs) for LASER (CCUCUUCUAAGCUCUUUAUTT), LSD1 (GAAGCTACATCTTACCTTA), EZH2 (GCTGAAGCCTCAATGTTTA), SREBP2 (CAAGGAGAGTCTATACTGT), CoREST(AAGAUUGUCCCGUUCUUGACU), LXRα (CTGCCCAGCAACAGTGTAA) and control scramble siRNA (UUCUCCGAACGUGUCACGUTT) were synthesized by GenePharma (Shanghai, China).
For RNAi assay, HepG2 cells were seeded at a density of 3.0 × 10⁵ cells per well in a 6-well plate, when the density of cell reach to 70–90% the next day, transfection was performed. After the density of the cells reached to 70–90%, transfection was performed by incubating the siRNA with Lipofectamine RNAiMAX in Opti-MEM for 30 min at room temperature to allow siRNA-lipid complexes to form. Then, the mixed reagents were added to the target cells for a final concentration of 50 nM siRNA. After incubation with siRNA for 48 hrs, the cells were harvested for the experiments.

siRNA targeting CCNB1 (CCNB1-siRNA, GAAUUCUGCACUAGUUCAA), Scramble siRNA control (Scramble-siRNA, GAUUAGUCACCUGACUAUA), miR-144 mimics (GGAUAUCAUCAUAUACUGUAAG), miR-144 inhibitor (CUUACAGUAUAUGAUGAUAUCC) and Scramble controls (GAGUACUACUUAUACAGUAUAG) were designed and synthesized by Shanghai GenePharma Company (Shanghai, China). Cells were seeded into 96-well plates in antibiotic-free growth medium at the density of 4 × 10³ cells per well. Transfection with was performed when the cells reached 70–80% confluence using Lipofectamine 2000 (Invitrogen, USA) in accordance to the manufacturer’s protocol.