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3 protocols using taurochenodeoxycholic acid

1

Baicalin and Bile Acid Regulation

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Baicalin (>98%) was purchased from Dalian Meilun Biotechnology Co., Ltd (Meilunbio, Dalian, China). EE was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Standards for BAs for cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), β-muricholic acid (β-MCA), taurocholic acid (TCA), taurodeoxycholic acid (TDCA), tauromuricholic acid (T-MCA), taurochenodeoxycholic acid (TCDCA), glycocholic acid (GCA), glycoursodesoxycholic acid (GUDCA), glycohyocholic acid (GHDCA), glycochenodeoxycholic acid (GCDCA), taurohyocholic acid (THDCA), glycodeoxycholic acid (GDCA), and interior label d4-glycochenodeoxycholic acid (d4-GCDCA) were obtained from Steraloids (Newport, RI, USA).
Antibodies directed against FXR, CAR, PXR, VDR, cholesterol 7a-hydroxylase (CYP7A1), and sterol 27-hydroxylase (CYP27A1) were purchased from Absin Biochemical Company (Shanghai, China). Antibodies directed against Sirt1, HNF-1α, tumor necrosis factor α (TNF-α), interleukin 1 beta (IL-1β), IL-6, and NF-κB were purchased from Cell Signaling Technology (Beverly, MA, USA). An antibody against BSEP was obtained from Santa Cruz Biotechnology (CA, USA). And an antibody against MRP2 was obtained from Abcam (Cambridge, UK).
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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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