MEFs were exposed to torin1 (475991, Merk Millipore, Burlington, MA, USA) and staurosporine (81590, Cayman Chemical, Ann Arbor, MI, USA), which had been pre-dissolved in dimethylsulfoxide (DMSO). MEF cells were seeded in Permanox 1-well Lab-Tek chamber slides from Nunc, Thermo Scientific (Rochester, New York, NY, USA) in Dulbecco’s Modified Eagle Medium (DMEM) (Life Technologies, Carlsbad, CA, USA) supplemented with 10% Fetal Bovine Serum (FBS) and 1% Penicillin-Streptomycin (Life Technologies, Carlsbad, CA, USA). Internal standards (IS) for hippuric acid-(phenyl-13C6), L-lysine-13C6−15N2, leucine-5,5,5-D3, D-glucose-13C6, glyceryl tri-(palmitate-1-13C), and cholic acid-2,2,4,4-D4 were purchased from Sigma-Aldrich (St. Louis, MI, USA). Caffeine-13C3 was acquired from Cerilliant Corporation (Round Rock, TX, USA), 18:1-D7 lyso-phosphatidylethanolamine (LPE), 18:1-D7 lyso-phosphatidylcholine (LPC) from Avanti Polar Lipids (Birmingham, UK), octanoyl-L-carnitine-(N-methyl-D3), ceramide [d18:1/18:1-(9Z)-13C18] and L-phenylalanine-13C9−15N from Cambridge Isotope Laboratories (Andover, MA, USA). Additional materials and chemicals are described in Supplementary Materials Section S1 .
Cholic acid 2 2 4 4 d4
Manufactured by Merck Group
Sourced in United States, Canada
Cholic acid-2,2,4,4-D4 is a deuterated cholic acid compound. It is a laboratory reagent used in research and analysis applications.
Automatically generated - may contain errors
Lab products found in correlation
Glyceryl tri palmitate 1 13c, by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
D glucose 13c6, by Merck Group (1 mentions)
13c6 15n2 l lysine, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Staurosporine, by Cayman Chemical (1 mentions)
N methyl n trimethylsilyltrifluoroacetamide mstfa, by Thermo Fisher Scientific (1 mentions)
Yeast extract, by Thermo Fisher Scientific (1 mentions)
Methoxyamine hydrochloride, by Merck Group (1 mentions)
Trimethylchlorosilane tmcs, by Thermo Fisher Scientific (1 mentions)
Hemin, by Merck Group (1 mentions)
L cysteine, by Merck Group (1 mentions)
Pyridine, by Merck Group (1 mentions)
Tryptone, by Thermo Fisher Scientific (1 mentions)
Resazurin sodium salt, by Merck Group (1 mentions)
2 3 5 triphenyltetrazolium chloride, by Merck Group (1 mentions)
2 ethylbutyric acid, by Merck Group (1 mentions)
3 protocols using cholic acid 2 2 4 4 d4
1
Metabolomic Analysis of MEF Cells
2
Extraction and Characterization of Crocin and Crocetin
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Crocin (PubChem CID: 5281233, purity: 99.33%) and crocetin (PubChem CID: 5281232, purity: 96.61%), Figure 1 , were purchased from Chengdu Biopurify Phytochemicals Ltd. (Chengdu, China). The Compound Edaravone Injection was provided by Jiangsu Simcere Pharmaceutical Co., Ltd. (Nanjing, China). 2,3,5-Triphenyltetrazolium chloride (TTC), stable-isotope-labeled IS 1,2-13C2-myristic acid, ES methyl stearate, methoxyamine hydrochloride, pyridine, hemin, resazurin sodium salt, L -cysteine, 2-ethylbutyric acid, and cholic acid-2,2,4,4-d4 were purchased from Sigma-Aldrich (St. Louis, United States). N-Methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) plus 1% trimethylchlorosilane (TMCS) was purchased from Thermo Scientific (Bellefonte, United States). Streptomycin sulfate and neomycin sulfate were purchased from Nanjing Xinhou Biological Technology Co., Ltd. (Nanjing, China). Yeast extract and tryptone were purchased from Oxoid (Basingstoke, United Kingdom).
3
Comprehensive Lipid Analytical Standards
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2.1. Materials and Chemicals. Solvents, modifiers, and their suppliers are described in the supporting information (SI; S1.1). 50 lipid analytical standards were purchased from Sigma Aldrich, Avanti Polar Lipids (Alabaster, USA), Cayman Chemical Company (Michigan, USA), and Honeywell Fluka (Charlotte, NC, USA) to cover a broad variety of human lipids originating from different categories including fatty acyls (15) (link), glycerolipids (5) (link), glycerophospholipids (11) (link), prenol lipids (1), sphingolipids (7) (link) and sterol lipids (11) (link). A detailed list of the lipid panel standards can be found in Table S1.2.1. Glyceryl tri(palmitate-1-13 C) and cholic acid-2,2,4,4-D4 were purchased from Sigma Aldrich, while lauric acid-12,12,12-D3 was bought from CDN Isotopes (Pointe-Claire, Quebec, Canada), 18:1-D7 lyso PE from Avanti Polar Lipids and octanoyl-L-carnitine-(N-methyl-D3) and ceramide (d18:1/18:1(9Z)-13 C18) from Cambridge Isotope Laboratories (Massachusetts, USA). For the in vitro experiments with liver cells, differentiated HepaRG cells were acquired from Biopredic International (Rennes, France).
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