E2F transcription factor-1 modulatesglutamine metabolism in transformed cells

To investigate the impact of E2F1 loss on glutamine metabolism we isolated mouse embryonic fibroblasts from E2f1-wildtype (wt) and E2f1-knockout (ko) mice We then performed gene expression profiling analysis using data obtained from RNA-seq of 2 different treatment conditions (standard growth media vs media containing glutamine but depleted of glucose ). Primary mouse embryonic fibroblasts (MEFs) were previously derived from E2f1-wt and E2f1-ko mice (B6;129S4- E2f1tm1 Meg/J) and SV40-immortalized. MEFs were cultivated with DMEM without glucose, glutamine, phenol red, and sodium pyruvate (A14430-01, Thermo Fisher Scientific) supplemented with 10% dialyzed and filtered FBS (26-400-044, Thermo Fisher Scientific), and either 25 mM glucose (G8769, Sigma-Aldrich) and 4 mM glutamine (25030-024, Thermo Fisher Scientific) or only 4 mM glutamine for 16 hours.