Dulbecco s modified eagle media

HCT116 p53+/+ and p53-/- (kindly provided by Prof. Bert Vogelstein, John Hopkins University, Baltimore, MD) isogenic colorectal carcinoma cell lines were used for the experiments [30 (link), 31 (link)]. HCT116 cells were grown in high glucose DMEM (Dulbecco’s Modified Eagle Media; Lonza) supplemented with 8 mM glutamine (Sigma-Aldrich), 1x antibiotic–antimycotic solution (Sigma-Aldrich), and 10% fetal bovine serum (FBS; Lonza). U2OS cells were maintained in low glucose DMEM (Dulbecco’s Modified Eagle Media; Lonza) supplemented with 4 mM glutamine (Sigma-Aldrich), 1x antibiotic–antimycotic solution (Sigma-Aldrich), and 10% fetal bovine serum (FBS; Lonza). Cells were maintained at 37°C in humidified atmosphere with 5% CO₂. Our cell culture-related study had been approved to be performed in the University of Szeged according to the TMF/43-18/2015 decision before the study began.

The murine dendritic cell line, JAWS II, was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). JAWS II was cultured in the presence of alpha-minimum essential media (Lonza, Basel, Switzerland) complemented with 20% fetal bovine serum (FBS, Life Technologies; Grand Island, NY, USA), 1% penicillin-streptomycin (Sigma-Aldrich, St. Quentin Fallavier, France), 4 mM L-glutamine (Lonza), 1 mM sodium pyruvate (Lonza), and 5 ng/mL of Granulocyte-Macrophage Colony Stimulating Factor (Miltenyi Biotec; San Diego, CA, USA). The cells were maintained in culture in a humified incubator at 37 °C and 5% CO₂.
The MDA-MB-231 human breast cancer cells were purchased from ATCC and cultured at 5% CO₂ at 37 °C in Dulbecco’s modified Eagle media (Lonza), supplemented with 10% FBS, 2 mM glutamine, 10% penicillin-streptomycin.
Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza and grown at 37 °C and 5% CO₂ in endothelial basal cell growth medium, supplemented with 2% FBS, 1% streptomycin-penicillin, 0.1% ascorbic acid, 0.1% human epidermal growth factor, 0.1% heparin, 0.1% VEGF, 0.1% gentamicin-amphotericin B, 0.04% hydrocortisone, 0.4% human bFGF, and 0.1% R3-IGF-1 (all from Lonza).

PC12 cells (PC-12 ATCC CRL-1721) were cultured and maintained in Dulbecco’s Modified Eagle Media (Lonza) supplemented with 10% heat inactivated fetal bovine serum (Sigma), 5% heat inactivated horse serum (Sigma) and 1% Penicillin/Streptomycin (Sigma). Cells were incubated at 37°C in a humidified incubator (5% CO₂). PC12 cells were pretreated for 1h with either: 0.1 μM [Hyp³]-Bradykinin (B7775, Sigma); 1 μM HOE-140 (BML-NK104-0001, Enzo Life Sciences); 0.1 μM Bradykinin Fragment 1–8 acetate salt hydrate (B4397, Sigma); 1 μM R-715 TFA salt (R9032, Sigma). All pre-treatments were reconstituted in dH₂O. Following pre-treatment, PC12 cells were treated for 3h, 12h and 24h with Staurosporine 1 μM (569397-100UG, Calbiochem), reconstituted in DMSO.