L glutamine

All tumor cell lines in this study were of metastatic origin. MDA-MB-231 and MCF-7 (both from American Type Culture Collection ATCC, Manassas, VA, USA) were cultivated in DMEM high glucose (Thermo Fisher Scientific, Waltham, MA, USA), 2 mM L-glutamine (Sigma-Aldrich, St. Louis, MO, USA), 10% FCS Superior (standardized fetal bovine serum, Biochrom GmbH, Berlin, Germany), and non-essential amino acids (Gibco Life Technologies, Carlsbad, CA, USA). Mel Im was cultivated in DMEM low-glucose (Sigma-Aldrich) with 2 mM L-glutamine, MV3 in DMEM high-glucose (Sigma-Aldrich) with 2 mM L-glutamine, with the addition of 10% FCS each. MV3 were obtained from Peter Friedl (RIMLS Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands). The media were supplemented with penicillin/streptomycin (100 U mL⁻¹, 0.1 mg mL⁻¹, Sigma-Aldrich), and the incubator was set to 5% CO₂, at 37 °C.

The oral fibroblasts were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) (Sigma, Dorset, United Kingdom), supplemented with 2% L-glutamine (Sigma, Dorset, United Kingdom), 100 IU/100 mg ml^-1 penicillin/streptomycin (Sigma, Dorset, United Kingdom), and 10% fetal calf serum (FCS) (Sigma, Dorset, United Kingdom).
The OKF6/TERT-2 human oral keratinocyte cells were cultured in Green’s medium, which consists of Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F12 medium in a 3:1 ratio, and is supplemented with 10% fetal calf serum (FCS), 10 ng/mL epidermal growth factor, 0.4 µg/mL hydrocortisone, 0.1 mM adenine, 5 µg/mL insulin, 5 µg/mL transferrin, 0.2 µM triiodothyronine, 2 mg/mL L-glutamine, 50 U/mL penicillin, and 50 U/mL streptomycin. All the agents were purchased from Sigma-Aldrich (Dorset, United Kingdom). Since live bacteria were not used in this study, the use of antibiotics in the cell culture medium would have minimal impact on the results.
The THP-1 monocytes were cultured in RPMI 1640 culture medium (Sigma, Dorset, United Kingdom) supplemented with 2 mM L-glutamine and 10% FCS.
Cultures were maintained in the incubators at 37 °C and 5% CO₂. The cells were cultured until 80–100% confluency was achieved.

Cells were cultured at 37°C and 5% CO₂. CH12F3 (22 (link)) cell clones were cultured in RPMI-1640 (Sigma) supplemented with 2 mM l-glutamine (Sigma), 10% FCS (heat-inactivated at 56°C for 30 min), 50 μM 2-β-mercaptoethanol (Gibco), and 1 × PenStrep solution (Gibco). U2OS cells were cultured in DMEM–high glucose (Sigma), 2 mM l-glutamine (Sigma), 10% FCS, 1 × PenStrep solution and 1.25 μg/ml Amphotericin B (Sigma). Wildtype, Ung^−/− (23 (link)), Aicda^−/− (24 (link)) mice were in C57BL6/J background (bred for >10 generations). The mice were housed at the IRCM specific pathogens-free animal facility. Mouse work was reviewed and approved by the animal protection committee at the Instituts de Recherches Cliniques de Montreal (protocol 2015–10). Naïve resting B cells were isolated from spleens from three nine to ten-month-old mice using EasySep™ Mouse B cell Isolation kit (STEMCELL) according to the manufacturers’ instructions. The isolated B cells were cultured in RPMI-1640 supplemented with 2 mM l-glutamine, 10% heat-inactivated FCS, 1 mM sodium pyruvate, 50 μM 2-mercaptoethanol, and 1 × PenStrep and stimulated with 10 μg/ml LPS (from E. coli strain 0111:B4, Merck) and 40 ng/ml IL-4 (PeproTech).

MV4-11 cells were cultured in the RPMI 1640 medium (HIIET PAS, Poland) supplemented with 1.0 mM sodium pyruvate, 2 mM L-glutamine and 10% FBS (all from Sigma Aldrich, Steinheim, Germany), LoVo cells were cultured in the F-12K medium (Life Technologies, Carlsbad, CA, United States) supplemented with 10% FBS (GE Healthcare, Chicago, IL, USA), and LoVo/DX cells were cultured in the mixture of RPMI 1640+OptiMEM (1:1) medium (HIIET PAS) supplemented with 5% FBS (GE Healthcare, Chicago, IL, USA), 2 mM L-glutamine, 1.0 mM sodium pyruvate (all from Sigma Aldrich, Steinheim, Germany) and 0.1 µg/mL doxorubicin chloride (Accord).
MCF-7 cells were cultured in the Eagle’s medium (HIIET PAS, Poland) supplemented with 10% FBS, 8 µg/mL insulin, 2 mM L-glutamine and 1% MEM-non essential amino acid solution 100X (all from Sigma Aldrich, Steinheim, Germany). MCF-10A cells were cultured in the HAM’S F-12 with L-glutamine medium (Corning) supplemented with 10% Hors Serum (Gibco), 10 µg/mL insulin, 0.5 µg/mL hydrocortisone, 20 ng/mL EGFh and 0.05 mg/mL Cholera Toxin from Vibrio cholerae (all from Sigma Aldrich, Steinheim, Germany). All culture mediums were supplemented with 100 units/mL penicillin (Polfa Tarchomin S.A. Poland), and 100 µg/mL streptomycin (Sigma Aldrich). All cell lines were grown at 37 °C with 5% CO₂ humidified atmosphere.

The ZR-75-1 (ATCC #CRL-1500 pg, RRID: CVCL_0588), T-47D (HTB-133, RRID: CVCL_0553), and MDA-MB-453 (HTB-131, RRID: CVCL_0418) breast cancer cell lines were obtained from American Type Cell Culture Collection (ATCC; Manassas, VA, USA), and the MFM-223 cell line was obtained from the DMSZ-German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany, RRID: CVCL_1408). All cell lines were routinely tested for mycoplasma infection and authenticity confirmed by short tandem repeat profiling (Cell Bank Australia). ZR-75-1 and T-47D cells were maintained in RPMI-1640 medium (Invitrogen) containing 10% Fetal Bovine Serum (FBS) and 2 nM L-Glutamine (Sigma). MFM-223 cells were cultured in EMEM (Sigma) containing 10% FBS, 2 nM L-Glutamine (Sigma), 1 × Non-essential Amino Acids (Sigma), and 1 × Insulin–Transferrin–Sodium Selenite (Sigma). MDA-MB-453 cells were maintained in DMEM (Sigma) medium containing 10% FBS, 2 nM L-Glutamine (Sigma) and 1 × Sodium Pyruvate (Sigma). All lines were incubated at 37 °C and 5% CO₂.

Human thyroid cancer cell lines, BCPAP harboring the BRAF V600E mutation and TPC-1 bearing the RET/PTC rearrangement, were a gift of Prof. M. Santoro (Medical School, University “Federico II” of Naples, Naples, Italy). These cell lines had been previously tested and authenticated by DNA analysis. Cancer cells were propagated in Dulbecco’s Modified Eagle Medium (DMEM) (Sigma, Saint Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma, Saint Louis, MO, USA), 2 mM L-glutamine and 100 U/ml penicillin/streptomycin (Sigma, Saint Louis, MO, USA) as previously described^{37 (link)}. Human thyroid cancer cell lines, 8305C and 8505C harboring the BRAF V600E mutation were previously tested and authenticated by DNA analysis. 8305C cells were propagated in Mininum Essential Medium (MEM) (Sigma, Saint Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma, Saint Louis, MO, USA), 2 mM L-glutamine and 100 U/ml penicillin/streptomycin (Sigma, Saint Louis, MO, USA). 8505C cells were propagated in RPMI medium (Sigma, Saint Louis, MO, USA) supplemented with 10% fetal bovine serum (Sigma, Saint Louis, MO, USA), 2 mM L-glutamine and 100 U/ml penicillin/streptomycin (Sigma, Saint Louis, MO, USA). Cells were incubated with the chosen stimuli in serum-free medium.