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3 protocols using α muricholic acid

1

Synthesis and Characterization of Bile Acid Derivatives

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Deoxycholic acid (DCA), chenoDeoxycholic acid (CDCA), cholic acid (CA), α-muricholic acid, β-muricholic acid, glycoDeoxycholic acid (GDCA), glycochenoDeoxycholic acid (GCDCA), and glycocholic acid (GCA) were obtained from Steraloids, Inc. Bile acid structures are shown in fig. S1. Bile acid A (37 ), 5-cholanic acid-3α,4β,7α-triol (7)(38 (link)), and AMPP (39 (link)) were synthesized according to published procedures. N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), 4-(dimethylamino)pyridine (DMAP), diethylamine, N,N-dimethylformamide, trisodium citrate, sodium hydroxide (NaOH), chloroform, Dulbecco’s modified Eagle’s medium, fetal calf serum, and penicillin G and streptomycin sulfate were obtained from Sigma–Aldrich. HPLC solvents (methanol and acetonitrile) were HPLC grade and were purchased from EMD Chemicals. Milli-Q ultrapure water was prepared in-house with a Milli-Q Integral Water Purification System.
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2

Bile Acid Compound Sourcing Protocol

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Taurocholic acid (TCA), tauro-β-muricholic acid (TMCA), tauromurideoxycholic acid (TMDCA), taurochenodeoxycholic acid (TCDCA), taurodeoxycholic acid (TDCA), taurolithocholic acid (TLCA), cholic acid (CA), α-muricholic acid (αMCA), β-muricholic acid (βMCA), ω-muricholic acid (ωMCA), murideoxycholic acid (MDCA), ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), and lithocholic acid (LCA) were purchased from either Steraloids, Inc. (Newport, RI) or Sigma-Aldrich (St Louis, MO). Clofibric acid (CLOF) was purchased from Sigma-Aldrich Co. (St. Louis, MO). Chloroform, agarose, and ethidium bromide were purchased from AMRESCO Inc. (Solon, OH). RNA-Bee was purchased from TelTest Inc. (Friendswood, TX). All other chemicals, unless indicated, were purchased from Sigma-Aldrich Co. (St. Louis, MO).
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3

Quantifying Bile Acid Profiles

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BA standards including cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), ursodeoxycholic acid (UDCA) and hyodeoxycholic acid (HDCA) were purchased from Sigma-Aldrich Chemicals Co (Steinheim, Germany). α-muricholic acid (α-MCA), β-muricholic acid (β-MCA), ω-muricholic acid (ω-MCA), tauro-cholic acid (T-CA), tauro-chenodeoxycholic acid (T-CDCA), tauro-deoxycholic acid (T-DCA), tauro-lithocholic acid (T-LCA), tauro-ursodeoxycholic acid (T-UDCA), tauro-α-muricholic acid (T-α-MCA), tauro- β-muricholic acid (T-β-MCA), glyco-cholic acid (G-CA), glyco-chenodeoxycholic acid (G-CDCA), glyco-deoxycholic acid (G-DCA), glyco-lithocholic acid (G-LCA), glyco-ursodeoxycholic acid (G-UDCA) and deuterated internal standards (IS) lithocholic acid-2,2,4,4-D4 (LCA-D4), deoxycholic acid-2,2,4,4-D4 (DCA-D4) and glycochenodeoxycholic acid-2,2,4,4-D4 (GCDCA-D4) were obtained from Steraloids, Inc. (Newport, RI, USA). Methanol HPLC grade was from Merck (Darmstadt, Germany). Ultra-pure reagent water was purified by a Milli-Q gradient system (Millipore, Bedford, MA, USA). Acetonitrile LC-MS grade was from VWR and formic acid LC-MS grade was purchased from Sigma Aldrich Chemicals Co.
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