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5 protocols using bms309403

1

Lipid Metabolism Modulation Protocol

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FBS was purchased from Life Technologies. Delipidized serum was purchased from Gemini Bio. CAY10566, BMS309403, sulfosuccinimidyl oleate, lipofermata, and etomoxir were purchased from Cayman Chemical. SC26196 was purchased from Santa Cruz. Avasimibe was purchased from Selleckchem. Glucose-D7, palmitic acid-D31, and oleic acid-D34 were purchased from Cambridge Isotope Laboratories. Palmitate, palmitoleate, oleate, cholesteryl oleate, and glyceryl trioleate were purchased from Sigma-Aldrich. Sapienate was purchased from Matreya. Octadecenoate (8(Z)-octadecenoic acid) was purchased from Larodan.
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2

Hypoxia-Induced Adipogenic Differentiation

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C3H10T1/2 (mouse fibroblast) cells were treated with HIF1 inhibitor (InSolution™ HIF‐1 inhibitor, Cat# 400092; Calbiochem, USA) at 10 or 20 μM. The cells were incubated in a hypoxic chamber for 48 h in the presence or absence of HIF1 inhibitor. Dimethyl sulfoxide (DMSO) was used as a vehicle. FABP4 inhibitor (BMS309403, Cat#10010206; Cayman, USA) was dissolved in the manufacturer‐recommended solvent, DMSO. One week after RCT, mice were treated daily with BMS309403 by oral gavage (15 mg/kg) for 2 weeks. Dimethyl sulfoxide was used as a vehicle. For hypoxic challenge, C3H10T1/2 cells were incubated in a hypoxic chamber (0.1% O2, 5% CO2, 10% H2, 85% N2) for the designated period.
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Comprehensive Cannabinoid Pharmacology Protocol

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The native phytocannabinoids Δ9-tetrahydrocannabinol (Δ9-THC, also called dronabinol, psychoactive high affinity CB1 and CB2 agonist) and cannabidiol (non-psychoactive, very low affinity CB1 and CB2 indirect antagonist thought to attenuate the action of Δ9-THC) were obtained from Cayman Chemical (Ann Arbor, MI). Synthetic cannabinoids obtained from Cayman Chemical (Ann Arbor, MI) were as follows: dronabinol (also called Δ9-tetrahydrocannabinol or Δ9-THC; high affinity CB1 and CB2 agonist), HU-210 (potent CB1 and CB2 receptor agonist analogue of Δ9-THC), JWH 018 (high affinity CB1 and CB2 ligand, mildly selective for CB2); Rimonabant high affinity selective CB1 receptor inverse agonist), JWH-133 (high affinity, 200-fold selective CB2 agonist), SR-144528 (high affinity CB2 inverse agonist), CP55,940 (high affinity non-selective CBR agonist, more potent than Δ9-THC). Synthetic cannabinoids that inhibit AEA uptake without affecting AEA hydrolysis (AM404), inhibit AEA uptake and weakly inhibit AEA hydrolysis (OMD1, OMD2), or inhibit AEA hydrolysis (VDM11) were purchased from Cayman Chemical (Ann Arbor, MI). BMS309403, known to bind/inhibit other FABPs (FABP3,4,5,7) was obtained from Cayman Chemical (Ann Arbor, MI). SCP2 inhibitors (SCPI1, SCPI3, and SCPI4) were from ChemBridge Corporation (San Diego, CA).
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4

Fatty Acid Isotope Labeling Protocol

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Glucose-d7, palmitic acid-d31 (PA-d31) and oleic acid-d34 (OA-d34) were purchased from Cambridge Isotope Laboratory. 17-Octadecynoic Acid (ODYA), BMS309403, cisplatin, and etomoxir were purchased from Cayman Chemicals. For treatment with cisplatin, 3.3 µM was used as the final concentration, unless otherwise specified.
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5

Fatty Acid Metabolism and Inhibition

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Chemicals were of the purest analytical grade. Bindarit was provided by Angelini Research Center/ACRAF (Rome, Italy). RPMI 1640, fetal bovine serum (FBS), and other cell culture reagents were purchased from Corning (Corning, NY, USA). BMS309403 was from Cayman Chemical (Ann Arbor, MI, USA), JX-401 (Abcam, Cambridge, UK) and CCT128930 (Selleck Chem, Munich, Germany). Oleic acid, [9,10-3H(N)] (37 MBq), arachidonic acid [5,6,8,9,11,12,14,15-3H] (1850 KBq), Lipidex 1000 resin and Ultima Gold XR were purchased from PerkinElmer (Milan, Italy). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), unless stated otherwise.
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