Mir 19a inhibitor

SH-SY5Y cells (ATCC) were incubated in Dulbecco’s modified Eagles’s medium (DMEM; Hyclone, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, USA) at 37°C in a humidified atmosphere with 5% CO₂. HEK-293T cells were cultured in RPMI-1640 medium (Gibco, Grand Island, NY, USA) supplied with 10% fetal bovine serum, 100U/ml penicillin, and 100 μg/ml streptomycin, at 37°C in a 5%CO₂ atmosphere.
For OGD model, the SH-SY5Y cells were incubated in serum and sugar-free artificial cerebrospinal fluid at 37°C in an atmosphere of 1% O₂, 94% N₂ and 5% CO₂ for 4 h and then incubated in DMEM medium in normoxic atmosphere.
In addition, miR-19a mimics, miR-19a inhibitor and their scramble controls were obtained from Guangzhou RiboBio Co., Ltd. (Guangzhou, China). The siRNAs (named as siRNA-1, siRNA-2 and siRNA-3) directed against H19 and negative control H19 scramble were purchased from Invitrogen Life Technologies (Waltham, MA, USA). The sequences of siRNAs were listed in Table 1.

HASM cells were transferred at a density of 2x10⁵ cells in 6-well plates and incubated for 24 h under the aforementioned conditions. The cells were transfected with 100 nM inhibitor negative control, 50 nM negative control mimics, 50 nM miR-19a mimics or 100 nM miR-19a inhibitor (RIBOBIO, Guangzhou, China) using lipofectamine 2000 reagent (Invitrogen, Carlsbad, USA) following the manufacturer’s protocol. Following 48 h of incubation, the expression levels of miR-19a were detected by Real-Time PCR.