3h uracil

The following tritium radiolabeled compounds were used during the project: [³H]-Uracil (40 Ci/mmol), [³H]-Thymidine (71.7 Ci/mmol), [³H]-Uridine (60 Ci/mmol) were obtained from PerkinElmer (Waltham, MA, USA). [³H]-Cytidine (20 Ci/mmol) was purchased from American Radiolabeled Chemicals Incorporated (St Louis, MO, USA). Uridine, Uracil, Thymidine, 5-FlourUracil, 5-FlouroUridine, 2’-DeoxyUridine, 3’-deoxyThymidine, Sulfadiazine, NBMPR, Adenosine, Inosine, resazurin sodium salt and Phenyl Arsine Oxide were bought from Sigma-Aldrich (Poole, UK). 5-Flouro 2’DeoxyUridine was from VWR; 5-MethylUridine was from Alfa Aesar; 2’,3’-dideoxyUridine was from Carbosynth; Pyrimethamine was from Fluka; Ara-A was from ICN Biomedicals.

All chemicals were obtained from Sigma (St Louis, MO) or Fisher (Waltham, MA) unless otherwise stated. [9,10-³H]oleic acid (sp act: 50 Ci/mmol) was purchased from Moravek (Brea, CA), [³H]uracil from PerkinElmer (Shelton, CT), 5-Butyl-4,4-Difluoro-4-Bora-3a,4a-Diaza-s-Indacene-3-Nonanoic Acid (C4-BODIPY-C9) from ThermoFisher Scientific (Waltham, MA) and 4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene (BODIPY 493/503) from Life Technologies (Carlsbad, CA). Non-polar lipid mixture for thin-layer chromatography was purchased from Matreya LLC (State College, PA). Atglistatin was purchased from Cayman Chemical (Ann Arbor, MI). The primary antibodies include: rabbit polyclonal anti-GRA7 [34 (link)], the rabbit polyclonal anti-aldolase and mouse monoclonal anti-Hsp70 (from Fidel Zavala, Johns Hopkins university, Baltimore), and the mouse monoclonal anti-SAG1 (from Jean-Francois Dubremetz, University of Montpellier, France). Secondary antibodies used for immunofluorescence were conjugated to Alexa⁴⁸⁸, Alexa⁵⁹⁴ or Alexa³⁵⁰ (Invitrogen, Carlsbad, CA). To prepare oleic acid (OA)-albumin complexes, sodium oleate was dissolved in H₂O at a concentration of 100 mM, then thoroughly mixed by vortexing for 3 min with 5% Fatty Acid Free BSA to reach a final concentration of 10 mM OA-BSA complexes.