Il 10 sirna

Human NP cells were purchased from American Type Culture Collection (ATCC) and cultured in Dulbecco's Modified Eagle Medium (DMEM)/F12 medium (with 10% fetal bovine serum and 1% penicillin‐streptomycin) at 37°C using a 5% CO₂ incubator. Cells were cultured in monolayer and the 2−3 generations of cells were used in the following experiments. Experiments were performed for three times.
To construct an in vitro model of IDD, we treated NP cells with 10 ng/ml of LPS for 24 h.
The lncRNA MAGI2‐AS3 sequence was synthesized based on the lncRNA MAGI2‐AS3 sequence and then subcloned into the pcDNA3.1 vector (MAGI2‐AS3‐plasmid; Shanghai GeneChem Co., Ltd.). The empty pcDNA3.1 vector was used as a control (control‐plasmid). For IL‐10 knockdown, the control‐siRNA (sc‐36869) and IL‐10‐siRNA (sc‐39634) were purchased from Santa Cruz Biotechnology. NP cells were transfected with control‐plasmid, MAGI2‐AS3‐plasmid, control‐siRNA, IL‐10‐siRNA, mimic control (Shanghai GenePharma), miR‐374‐5p mimic (Shanghai GenePharma), inhibitor control (Shanghai GenePharma), miR‐374‐5p inhibitor (Shanghai GenePharma), MAGI2‐AS3‐plasmid+mimic control, MAGI2‐AS3‐plasmid + miR‐374‐5p mimic, miR‐374‐5p inhibitor+control‐siRNA, or miR‐374‐5p inhibitor + IL‐10‐siRNA using Lipofectamine 2000 reagent (Thermo Fisher Scientific, Inc.) at 37°C for 24 h as per the manufacturer's protocol.