Human colon adenocarcinoma LS174T cells (ATCC, Manassas, VA) were cultured in 24-well plates with phenol red-free Minimum Essential Media (Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum at 37°C in an atmosphere containing 5% CO2. On day 4, cells were transiently transfected with the pCMV-ICIS human NR1H4/FXR plasmid (LS174T-FXR, Open Biosystems, Huntsville, AL) (Li et al., 2012) using Lipofectamine LTX according to the manufacturer’s recommendations (ThermoFisher Scientific, Rockford, IL). On day 5, cells were cultured in media supplemented with 10% charcoal stripped fetal bovine serum and treated with vehicle (0.1% DMSO), 0.1 μM 17β-estradiol, 1 μM progesterone, 7 μM epiallopregnanolone-sulfate (PM5S, Steraloids, Newport, RI), 0.3 μM placental lactogen (NHPP, Torrance, CA), 13.3 nM testosterone, 7 pM growth hormone, 0.8 μM cortisol or 50 nM dehydroepiandrosterone (Steraloids), concentrations that represent third trimester levels in humans (Abu-Hayyeh et al., 2010 (link); Jeong, 2010 (link)). Total RNA was collected from cells on day 6.
Dehydroepiandrosterone
Manufactured by Steraloids
Sourced in United States
Dehydroepiandrosterone is a naturally occurring steroid hormone. It is the most abundant circulating steroid in the human body and serves as a precursor for the production of other steroid hormones, including testosterone and estrogen. This product is intended for use in research and laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Testosterone, by Steraloids (2 mentions)
Cortisol, by Steraloids (1 mentions)
Epiallopregnanolone sulfate pm5s, by Steraloids (1 mentions)
Ls174t cells, by American Type Culture Collection (1 mentions)
Minimum essential media, by Thermo Fisher Scientific (1 mentions)
Lipofectamine ltx, by Thermo Fisher Scientific (1 mentions)
Androsterone, by Steraloids (1 mentions)
Etiocholanolone, by Steraloids (1 mentions)
Epitestosterone, by Steraloids (1 mentions)
Testosterone d3, by Steraloids (1 mentions)
Pregnenolone, by Steraloids (1 mentions)
Progesterone, by Steraloids (1 mentions)
Diethyl ether, by Thermo Fisher Scientific (1 mentions)
Finasteride, by Merck Group (1 mentions)
Hplc grade methylene chloride, by Thermo Fisher Scientific (1 mentions)
3 protocols using dehydroepiandrosterone
1
Transfection and Treatment of LS174T Cells
2
Steroid Quantification Protocol
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Testosterone, epiTestosterone, androsterone, etiocholanolone, 5α-androstane-3α,17β-diol, 5βandrostane-3α,17β-diol, dehydroepiandrosterone, 5β-androstanedione, 4-androstenedione and Testosterone-d3 were purchased from Steraloids Inc. (Newport, RI, USA); 5α-dihydroTestosterone was provided by LGC Promochem SRL (Milan, Italy). Isopropyl alcohol, methanol, methyl tertbutyl ether (TBME), ethyl acetate, 17α-methylTestosterone, dithioerythritol and N-Methyl-N-
3
Steroid Acquisition and Preparation
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Steroids (progesterone, pregnenolone, testosterone, dehydroepiandrosterone (DHEA) and DHP) were purchased from Steraloids (Newport, RI, USA). Enzyme inhibitors were purchased from the following sources: trilostane, Sanofi Research Division, Great Valley, PA, USA; finasteride, Sigma Chemical Co, St Louis, MO, USA; dutasteride, Aurum Pharmatech, Howell, NJ, USA. Methylene chloride (HPLC grade) and diethyl ether were purchased from Fisher (Fair Lawn, NJ, USA) and ethyl acetate (chromatography grade) from EMD Millipore.
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