Mcdb 131 cell culture medium

MCDB-131 cell culture medium was obtained from Gibco (Paisley, Scotland, UK). Other cell culture media, penicillin, streptomycin and amphotericin B, and trypsin were purchased from BioWhittaker (Verviers, Belgium). Fetal bovine serum (FBS) was purchased from Biowest (Kansas, USA). Matrigel was purchased from Becton-Dickinson (Bedford, MA, USA). Fasentin was supplied by Calbiochem (San Diego, CA, USA), dissolved in DMSO as a 70 mM stock and stored at −20 °C. In all the assays, the vehicle (DMSO) was lower than 0.14% (v/v). STF-31 was supplied by Sigma-Aldrich (St. Louis, MO, USA), dissolved in DMSO as a 40 mM stock and stored at −20 °C. 2-NBDG was supplied by Molecular Probes (Eugene, OR, USA). All the antibodies used in this work were from Cell Signaling Technology (Danvers, MA, USA). Fertilized chick eggs were obtained from Granja Santa Isabel (Córdoba, Spain). Wild-type AB zebrafish were kindly supplied by Dr. Manuel Marí Beffa (Universidad de Málaga, Spain). Plastic material for cell culture was from Nunc (Roskilde, Denmark). All other reagents not listed on this section were from Sigma-Aldrich (St. Louis, MO, USA).

MCDB-131 cell culture medium was obtained from Gibco (Paisley, Scotland, UK). Other cell media, penicillin, streptomycin and amphotericin B, and trypsin were purchased from BioWhittaker (Verviers, Belgium). Fetal bovine serum (FBS) was purchased from Biowest (Kansas, USA). 2-NBDG (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose) and BODIPY FL C₁₆ (4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-hexadecanoic acid) were supplied by Molecular Probes (Eugene, OR, USA). L-[¹⁴C(U)]-Glutamine was acquired from Perkin Elmer (Waltham, Massachusetts, USA). Lactate assay kit was from Abnova (Taoyuan City, Taiwan). Anti-MCT1 antibody was from Santa Cruz Biotechnology (Heidelberg, Germany), anti-HIF-1α antibody was from BD Biosciences (Franklin Lakes, NJ, USA) and anti-LC3B and anti-α-tubulin antibodies were from Cell Signaling Technology (Danvers, MA, USA). Plastic material for cell culture was from Nunc (Roskilde, Denmark). All other reagents not listed on this section, including sodium palmitate, were from Sigma-Aldrich (St. Louis, MO, USA).