Cell Culture of Glioblastoma and Fibroblast Lines

NIH-3T3 mouse embryo fibroblasts, A172 human glioblastoma, U251 human glioblastoma, U87MG human glioblastoma, RKO human colon carcinoma, and HT-1080 human fibrosarcoma cells were obtained from American Type Culture Collection (Manassas, VA, USA). The generation of stable p53 knockdown A172 cell lines using lentivirus has been previously described (Gorjala et al., 2016 (link)). These cells express shRNA that knocks down p53 expression, or a non-targeted shRNA generated in parallel that is used as a control. All cells were grown in RPMI 1640 supplemented with 10% fetal bovine serum (10% calf serum for NIH-3T3 cells), 25 mM HEPES and 1× antibiotic-antimycotic (Invitrogen-Gibco) at 37°C in 5% CO₂.