Endogro mv vegf complete culture media kit

The human Sk-Hep1 cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA, catalog n° HTB-52) and was propagated in Dulbecco’s modified Eagle’s medium (DMEM; GIBCO by Thermo Fisher Scientific, Waltham, MA, USA) with 10% FBS (GIBCO), 100 U/mL penicillin, and 100 μg/mL streptomycin (GIBCO). Sk-Hep1 cells were used up to passages 20–25.
Human breast tumor-derived endothelial cells (BTECs) were isolated and characterized in the laboratory of Professor Benedetta Bussolati, Department of Molecular Biotechnology and Health Sciences, University of Torino, Italy [15 (link),16 (link)]. BTECs were maintained in EndoGRO-MV-VEGF Complete Culture Media Kit (Merck, Darmstadt, Germany, catalog n° SCME003), supplemented with antibiotics (100 U/mL penicillin and 100 ug/mL streptomycin; Gibco by Thermo Fisher Scientific, catalog n° 15140122).
Cells were maintained in a 37 °C and 5% CO₂ air incubator and routinely screened for absence of mycoplasma contamination.
To inhibit heme synthesis, FLVCR1a-overexpressing Sk-Hep1 cells were treated for 24h with 5 mM 5-aminolevulinic acid hydrochloride (Sigma Aldrich, St. Louis, MO, USA, catalog n° A3785). To inhibit citrate export from mitochondria to cytosol, FLVCR1a-silenced Sk-Hep1 cells were treated for 1h with 5 µM iCIC (Sigma Aldrich, catalog n° SML0068).