Gbm8401 glioma cell line

We extracted total mRNA via TRIzol™ Reagent (Thermo Fisher Scienti c, Wal-tham, WA, USA) according to the manufacturer's protocol. Oligo dT primer with MMLV Reverse Transcriptase (Epicentre Biotechnologies, Madison, WI, USA) was applied for cDNA synthesis. We purchased normal brain cDNA from Origene Technologies (Rockville, MD, USA). Ampli cation and quanti cation of DPY19L1 expression were achieved by a StepOne™ Real-Time PCR System (Thermo Fisher Scienti c, USA). We used the 2 -ΔΔCt method to compare relative quantitative gene expression with GAPDH as an internal control. The primer pairs included presented below: DPY19L1 forward 5′-ACACCACCTCTCCGTGAAAGCT-3' and reverse 5′-GCAGAGTGCAATCAA-GCTTCCTC-3'; GADPH forward 5′-CTTCATTGACCTCAACTAC-3′ and reverse 5′-GCCATCCACAGTCTTCTG-3′.
Cell Culture LN229, U118MG and U87MG cell lines were commercially available from American Type Culture Collection (ATCC), and we also purchased GBM8401 glioma cell line from Bioresource Collection and Research Center (BCRC number 60163, Hsinchu, Taiwan). LN229 and GBM8401 cells were cultured in Dulbecco's modi ed Eagle's medium (DMEM) which comprises 2% fetal bovine serum (FBS), penicillin, and streptomycin, and U87MG, U118MG, and LNZ308 cells were cultured in DMEM consisting of 10% FBS, penicillin, and streptomycin. Cells above were incubated in 37°C and 5% CO2 condition.