Mimics nc

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.

Lentiviruses containing vector, MIR22HG overexpressing plasmid, negative control shRNA (shNC), shMIR22HG, shPrrx-2, inhibitor NC, miR-128 inhibitor, mimics NC, miR-128 mimics, YAP1 overexpressing plasmid were provided by GeneChem (Shanghai, China). BMSCs were infected with the above lentiviruses at a multiplicity of infection in the presence of 5 µg/mL polybrene according to a previous study (Yang et al. 2021 (link)). Transfection was carried out by using Lipofectamine 3000 (Invitrogen, Missouri, USA) according to manufacturer’s protocol. Cells were collected for further analysis 48 h after transfection. The stably transfected cells were selected using G418.

The shRNAs targeting SNHG14 or Mef2c, miR-493-5p inhibitor, miR-493-5p mimics, and their negative control (shNC, inhibitor NC, mimics NC) were synthesized by GeneChem (Shanghai, China). And cells were transfected with above plasmids by liposome 2000 reagent (Invitrogen, CA, USA). Then, lentiviral carried with overexpression vectors (OE-SNHG14) were co-transfected into human embryonic kidney 293 T (HEK293T) cells, and the three packaging plasmids included pHBLVTM, psPAX2, and pMD2.G. Lentiviral particles were obtained after 48 h of transfection via cell harvest and concentration with ultracentrifugation at 72,000 g for 2 h. Following titer determination, BMSCs were transfected at a multiplicity of infection (MOI) of 50 for 24 h.