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.
Epiallopregnanolone sulfate pm5s
Manufactured by Steraloids
Sourced in United States
Epiallopregnanolone-sulfate (PM5S) is a steroid compound used in research applications. It is a sulfated derivative of the neurosteroid allopregnanolone. PM5S is primarily utilized in scientific studies and laboratory experiments, but its detailed function and intended uses should not be extrapolated upon in this factual description.
Automatically generated - may contain errors
Lab products found in correlation
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Methanol, by Thermo Fisher Scientific (2 mentions)
Cortisol, by Steraloids (1 mentions)
Dehydroepiandrosterone, 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)
Isopropanol, by Thermo Fisher Scientific (1 mentions)
Progesterone, by Steraloids (1 mentions)
Reverse transcription kit, by Takara Bio (1 mentions)
D9 progesterone, by Steraloids (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Sybr green kit, by Takara Bio (1 mentions)
Rnaiso plus reagent, by Takara Bio (1 mentions)
2 protocols using epiallopregnanolone sulfate pm5s
1
Transfection and Treatment of LS174T Cells
2
Steroid hormone quantification protocol
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Methanol, acetonitrile, isopropanol, methanoic acid, fetal bovine serum, and DMEM medium were purchased from Thermo Fisher (Fairlawn, NJ, USA). Progesterone, Progesterone-d9, allopregnanolone sulfate (PM4S), and epiallopregnanolone sulfate (PM5S) were purchased from Steraloids (Newport, RI, USA). RNAiso Plus reagent, Reverse transcription kit, and SYBR Green kit were purchased from Takara (Dalian, China). The estradiol radioimmunoassay KIT was received from the Beijing North Institute of Biological Technology (Beijing, China). Charcoal, dextran coated, and other biochemical regents were obtained from Sigma (St. Louis, MO, USA), unless otherwise stated.
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