Mda mb 468 breast cancer cell line

The MDA-MB-468 breast cancer cell line used in this study was originally obtained from the American Type Culture Collection (Manassas, VA, USA) by the Georgetown Lombardi Comprehensive Cancer Center (Washington, DC, USA) [62 (link)]. Cell cultures were routinely maintained in DMEM (Sigma-Aldrich, St. Louis, MO, USA) containing glucose (4500 mg/L), l-glutamine, and sodium pyruvate (110 mg/L) and supplemented with 10% FBS (SAFC Biosciences, Castle Hill, Australia). Cultures were maintained in antibiotic-free growth medium at 37 °C in a humidified incubator with O₂ and CO₂ levels set at 21% and 5%, respectively. For induction of hypoxia (HPX) in culture, the cells were passaged in DMEM supplemented with 10% FBS and incubated at 37 °C on an InvivO₂ 400 workstation (Ruskinn Technology Ltd, Bridgend, UK) at 1% O₂ and 5% CO₂, and the cells were harvested at 48 h.

All human Burkitt lymphoma cell lines were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ). MDA-MB-468 breast cancer cell line was purchased from American Type Culture Collection (ATCC). BL cell lines were grown in RPMI-1640 medium supplemented with heat-inactivated 10% or 20% (RAMOS) fetal bovine serum (FBS), 100 U/mL penicillin, 100 U/mL streptomycin, and 25 mmol/L HEPES buffer (all from Lonza) at a density of 0.5–2.0 × 10⁶ cells/mL. MDA-MB-468 cell line was cultured in DMEM low glucose medium (Lonza) supplemented with FBS, penicillin, and streptomycin as above. All cells were grown in a humidified atmosphere at 37 °C with 5% CO₂.
PHGDH inhibitor NCT-503 (N-(4,6-dimethylpyridin-2-yl)-4-(4-(trifluoromethyl)benzyl) piperazine-1-carbothioamide) and NCT-503 inactive control (N-(4,6-dimethylpyridin-2-yl)-4-(pyridin-4-yl)piperazine-1-carbothioamide), unable to inhibit PHGDH) were purchased from Sigma-Aldrich. Glutaminase inhibitor CB-839 was purchased from MedChemExpress. All compounds were diluted in sterile dimethyl sulfoxide (DMSO) to obtain 10 mM stocks, aliquoted, and stored in 2–8 °C according to manufacturer’s recommendations.