RNA-Seq data from human GBM stem-like cells expressing transgenice TGFBR2
The goal of this experiment was to determine genes regulated by TGFBR2 in glioma stem-like cell. GBM1A (1A) and GBM1B (1B) were transduced with lentivirus to generate stable transgenic lines expressing TGFBR2 (1A-R2 and 1B-R2). Briefly, HEK293FT were co-transfected with second-generation lentiviral packaging system (psPAX2 and pMD2.G) from Addgene using the Lipofectamine 3000 kit (Thermo Fisher) according to the manufacturer's recommendations. Sodium butyrate (Cayman Chemical) was added to transfected cells at a final concentration of 10mM 24 hrs after transfection and lentiviral particles were collected, filtered, and concentrated using Lenti-X concentrator solution (Takara Bio) 48 hrs after cell transfection. Lentiviral particles were resuspended in 1mL PBS after concentration and added to neurospheres in the presence of 1ug/mL of polybrene (Santa Cruz Biotechnologies). The next morning, cells were replated in fresh neurosphere media. Plasmids for TGFBR2 (EX-Z4152-Lv242) were purchased from Gencopoeia.