Pcdna3.1 carrier

MKN-45, MGC803, AGS human gastric cancer cell lines and HEK-293T cell line were purchased from Procell Life Science&Technology Co., Ltd (Wuhan, China) in Jan 2019. All cell lines underwent short tandem repeats (STR)-authentication and mycoplasma contamination tests. And all cells were cultured with Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS) (Hyclone, Logan City, UT, USA) and 1% penicillin/streptomycin (Hyclone, Logan City, UT, USA) in a humidified atmosphere of 5% CO2 at 37 °C. We performed rapid freezing cryopreservation for first two to four generations of cells with 1mL freezing medium of 90% FBS and 10% Dimethyl sulfoxide (DMSO) in liquid nitrogen.
SiRNA against lncRNA HOXA11-AS (si-HOXA11-AS), negative control siRNA of random sequence (siRNA-NC), miR-124-3p mimics, negative control miR-124-3p mimics (mimics NC), miR-124-3p inhibitor (ASO-miR-124-3p), and negative control miR-124-3p inhibitor (ASO-NC) were all purchased from Genechem Co., Ltd, (Shanghai, China). HOXA11-AS cDNA sequences were amplified and cloned to pcDNA3.1 carrier (Invitrogen, Carlsbad, CA, USA). According to the manufacturer’s instructions of Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), cells transfection was conducted in six-well plates.

Dulbecco’s Modified Eagles Medium (DMEM), fetal bovine serum, and Opti-MEM were all purchased from Gibco of the United States. miR-9 mimic and inhibitor were purchased from Shanghai GenePharma Co.,Ltd. pcDNA3.1 carrier and Lipofectamine 2000 were purchased from Invitrogen of the United States. Methylthiazolyl tetrazolium (MTT) kit was purchased from Sigma of the United States. SYBR green PCR kit and cDNA reverse transcription kit were all purchased from Takara of Japan. Dimethyl sulfoxide (DMSO), bicinchoninic acid (BCA) protein detection kit, and poly (vinylidene fluoride) (PVDF) were all purchased from Shanghai Beyotime Biotechnology. ECL luminescent detection kit was purchased from Thermo fisher of the United States. Both protein primary antibody and secondary antibody were purchased from Abcam of the UK.