L glutamine

BSC-1 cells (ATCC CCL-26) were grown in Eagle’s Minimal Essential Medium (EMEM) supplemented with 0.1 mg/ml penicillin, 0.1 mg/ml streptomycin, 2 mM L-glutamine (Lonza) and 5% foetal bovine serum (FBS).
Flp-In T-Rex 293 cells (Invitrogen) were maintained in DMEM supplemented with 0.1 mg/ml penicillin, 0.1 mg/ml streptomycin, 2 mM L-glutamine (BioWhittaker), 7% foetal bovine serum (FBS), 300 µg/ml Zeocin and 10 µg/ml blasticidin.
The 293-Cre cell line, inducibly expressing Cre recombinase, was derived from Flp-In-293 cells by Flp recombinase-mediated insertion of plasmid pcDNA-FRT-Cre using the Flp-In T-Rex system (Invitrogen) following manufacturer’s instructions. 293-Cre cells were grown in DMEM supplemented with 7% foetal bovine serum (FBS) containing 0.1 mg/ml penicillin, 0.1 mg/ml streptomycin, 2 mM L-glutamine (BioWhittaker), 10 µg/ml blasticidin and 100 µg/ml hygromycin. Routinely, for induction of Cre expression, cells were incubated with medium containing 1 µg/ml tetracycline for 18–24 h before infection. After virus adsorption, incubation was continued in medium containing 2% FBS and 1 µg/ml tetracycline.

Jurkat cells were cultured in RPMI-1640 medium (Lonza, Basel, Switzerland) containing 10% heat-inactivated fetal bovine serum (FBS, Hyclone, Logan, UT, USA), 2 mM l-glutamine (Lonza), and 100 U/ml penicillin/streptomycin (Lonza).
HeLa cells were cultured in DMEM medium (Lonza) containing 10% heat-inactivated fetal bovine serum (FBS, Hyclone), 2 mM l-glutamine (Lonza), 100 U/ml penicillin/streptomycin (Lonza), 1 mM sodium pyruvate, and 0.1 mM nonessential amino acids (NEAA, Lonza).
Mouse CD4+ T cells were cultured in RPMI-1640 medium (Lonza) containing 7.5% heat-inactivated fetal bovine serum (FBS, Hyclone), 2 mM l-glutamine (Lonza), 100 U/ml penicillin/streptomycin (Lonza), and 50 μM β-mercaptoethanol (Sigma-Aldrich, St. Louis, MO, USA). All cells were incubated at 37°C in a 5% CO₂ environment.

U87MG (GFP or FABP3-GFP) cells were maintained with low glucose (1.0 g/L glucose) DMEM medium (Lonza, Walkersville, MD, USA) supplemented with 2 mM l-glutamine (Lonza), 1% penicillin/streptomycin (100 Units/mL penicillin and 100 µg/mL streptomycin, Lonza), and 10% fetal bovine serum (FBS, Biowest). BT12-GFP cells were maintained with the Dulbecco’s Modified Eagle’s Medium (DMEM)/F-12 (1:1) growth medium (Gibco) supplemented with 2 mM l-glutamine (Lonza), 1% penicillin/streptomycin (100 Units/mL penicillin and 100 µg/mL streptomycin, Lonza), 15 mM HEPES buffer (Lonza), 2% B27 (Gibco), 0.01 µg/mL recombinant human fibroblast growth factor (FBF-b, Peprotech), 0.02 µg/mL recombinant human epidermal growth factor (EGF, Peprotech), and 1 µg/mL doxycycline. 293FT cells were maintained with high glucose (4.5 g/L glucose) DMEM medium (Lonza) supplemented with 2 mM l-glutamine, 1% penicillin/streptomycin (100 Units/mL penicillin and 100 µg/mL streptomycin), and 10% fetal bovine serum (FBS). All cells were cultured in the humidified incubator at 37 °C under a 5% CO₂ atmosphere.

B16-F10 (ATCC; RRID:CVCL_0159), Hep-55.1 C (CLS; RRID: CVCL_5766), Hepa1-6 (ATCC; RRID: CVCL_0327), RIL175 (Prof. Greten), Pan02 (Prof. Lauber) and T110299 (Prof. Siveke; Ptf1a-Cre LSL-K_ras^G12D LSL-Trp53^fl/R172H) cells were cultured in DMEM (Lonza, Cologne, Germany) supplemented with 10% fetal calf serum (FCS; ThermoFisher Scientific, Darmstadt, Germany), 2 mM L-glutamine (Lonza, Cologne, Germany) and 10 µg/ml ciprofloxacin (Fresenius Kabi, Bad Homburg, Germany). B3Z T cell hybridoma cells were cultured in RPMI 1640 (Lonza, Cologne, Germany) supplemented with 10% FCS, 2 mM L-glutamine and 10 µg/ml ciprofloxacin. Splenocytes were cultured in RPMI 1640 supplemented with 10% FCS, 2 mM L-glutamine, 100 U/L penicillin, 0.1 mg/ml streptomycin (all Lonza, Cologne, Germany), 1 mM sodium pyruvate (Biochrom, Berlin, Germany) and 0.1 mM non-essential amino acids (ThermoFisher, Darmstadt, Germany). For T cell activation, cells were seeded at 1 × 10⁶ cells/0.5 ml in a 48-well plate, in the presence or absence of anti-CD3/anti-CD28 Dynabeads^® (ThermoFisher, Darmstadt, Germany). Cells remained in culture for 48 h and were subsequently processed for flow cytometry. For cell death induction, cells were treated with 1 µM staurosporine (Sigma Aldrich, Munich, Germany) for 4–6 hours.

HER2 positive breast adenocarcinoma cells (SKBR-3), ERα positive breast adenocarcinoma cells (MCF-7) used as cancer negative control, murine fibroblasts cells (NIH-3T3) used as negative control. All cell lines were obtained from the American Type Culture Collection (HTB85, ATCC, Manassas, VA, United States). After thawing, SKBR-3 cell line was cultured in McCoy’s 5A Medium Modified (Sigma-Aldrich), 10% of Fetal Bovine Serum (FBS, EuroClone), 1% of L-Glutamine (Lonza), 2% sodium pyruvate (Lonza), 0.4% antibiotics (Lonza) and 0.1% fungizone. MCF-7 cell line was cultured in Minimum Essential Medium Eagle (Sigma-Aldrich), 10% of Fetal Bovine Serum (EuroClone), 1% sodium pyruvate (Lonza), 1% Non-Essential Aminoacids (EuroClone), 1% Bovine Insulin (Sigma-Aldrich), 1% L-Glutamine (Lonza) and 0.4% antibiotics (Lonza). NIH-3T3 cell line was cultured in Dulbecco’s Modified Eagle Medium High Glucose 4.5 mg/mL (Sigma-Aldrich), 10% of Bovine Calf Serum (BCS, EuroClone) and 1% of L-Glutamine (Lonza). All cell lines were incubated at 37°C in 5% CO₂, routinely trypsinized (Trypsin EDTA solution 1X, Lonza) after confluence, counted and seeded into wells.

Cancerous cell lines (MCF-7: human breast adenocarcinoma cells; HepG2: human hepatocellular carcinoma cells; Caco-2: human epithelial colorectal adenocarcinoma cells; A549: human epithelial lung adenocarcinoma cells), obtained from the American Type Culture Collection (ATCC) (Rockville, MD, United States of America), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) high glucose (4.5 g/L) containing L-glutamine (4 mM) and sodium-pyruvate (Hyclone™) supplemented with 10% (v/v) fetal bovine serum (FBS) (Capricorn Scientific GmbH, South America), and incubated at 37°C with 5% CO₂ in a humidified environment. African green monkey (Vero) kidney cells (also obtained from ATCC), a non-cancerous cell line, were grown in DMEM high glucose (4.5 g/L) containing L-glutamine (Lonza, Belgium) and supplemented with 5% FBS (Capricorn Scientific GmbH, South America) and 1% gentamicin (Virbac, RSA) in the same environment as cancer cells. The RAW 264.7 murine macrophage cells (obtained from ATCC) were cultured in DMEM high glucose (4.5 g/L) containing L-glutamine (Lonza, Belgium), supplemented with 10% FBS (Capricorn Scientific GmbH, South America) and 1% penicillin/streptomycin/fungizone (PSF) solution, and kept at 37°C in a 5% CO₂ humidified environment.