Journal: Cell

Article Title: FXR regulates intestinal cancer stem cell proliferation

doi: 10.1016/j.cell.2019.01.036

Figure Lengend Snippet: WT and APCmin/+ mice on ND and HFD were treated as described in Figure 5A and ​andB.B. Experimental schemes of Vehicle (black) and FexD-treated (50mg/kg/day p.o., yellow) WT and Vehicle (red) and FexD-treated (green) APCmin/+ mice as described in Figure 5A, ​,CC.

Article Snippet: Cell viability assay and Cell Luciferase assay FexD, OCA (Obeticholic acid), CDCA (Chenodeoxycholic acid) and GW4064 were dissolved in DMSO (FexD, in house production; OCA, GW4064, Selleck Chemical LLC).

Techniques:

Journal: Cell

Article Title: FXR regulates intestinal cancer stem cell proliferation

doi: 10.1016/j.cell.2019.01.036

Figure Lengend Snippet: (A) Brightfield images of organoids generated from APCmin/+ mice on ND, treated from days 3–6 with vehicle (DMSO), T-βMCA, FexD, and T-βMCA+ FexD. Scale bar 50μm.

Article Snippet: Cell viability assay and Cell Luciferase assay FexD, OCA (Obeticholic acid), CDCA (Chenodeoxycholic acid) and GW4064 were dissolved in DMSO (FexD, in house production; OCA, GW4064, Selleck Chemical LLC).

Techniques: Generated

Journal: Cell

Article Title: FXR regulates intestinal cancer stem cell proliferation

doi: 10.1016/j.cell.2019.01.036

Figure Lengend Snippet: (A, B) Schematics of FexD treatment (50mg/kg/day p.o.) of WT and APCmin/+ mice on ND and HFD. Mice were fed HFD from 4 weeks.

Article Snippet: Cell viability assay and Cell Luciferase assay FexD, OCA (Obeticholic acid), CDCA (Chenodeoxycholic acid) and GW4064 were dissolved in DMSO (FexD, in house production; OCA, GW4064, Selleck Chemical LLC).

Techniques:

Journal: Pharmaceutics

Article Title: Toxicokinetics of β-Amanitin in Mice and In Vitro Drug–Drug Interaction Potential

doi: 10.3390/pharmaceutics14040774

Figure Lengend Snippet: Inhibition of UGT1A1-catalyzed SN-38 glucuronidation, UGT1A3-catalyzed chenodeoxycholic acid 24-acyl-glucuronidation, UGT1A4-catalyzed trifluoperazine N -glucuronidation, UGT1A6-catalyzed N -acetylserotonin glucuronidation, UGT1A9-catalyzed mycophenolic acid glucuronidation, and UGT2B7-catalyzed naloxone 3-β-D-glucuronidation by β-amanitin in human liver microsomes using cocktail substrates and LC-MS/MS. Each data point represents mean ± SD ( n = 3).

Article Snippet: N -acetylserotonin β-D-glucuronide, amodiaquine, bupropion, chenodeoxycholic acid 24-acyl-β-glucuronide, N -deethylamodiaquine, diclofenac, 4′-hydroxybupropion, 7-hydroxycoumarin, 4′-hydroxymephenytoin, mycophenolic acid β-D-glucuronide, propofol-β-D-glucuronide, SN-38 glucuronide, and [ S ]-mephenytoin were obtained from Toronto Research Chemicals Inc. (Toronto, ON, Canada).

Techniques: Inhibition, Liquid Chromatography with Mass Spectroscopy

Journal: Nature medicine

Article Title: Gut microbiota and intestinal FXR mediate the clinical benefits of metformin

doi: 10.1038/s41591-018-0222-4

Figure Lengend Snippet: a, TR-FRET FXR coactivator recruitment assay to assess whether GUDCA is a FXR antagonist in the presence of CDCA (20 μM). n = 4 replicates/treatment. b, Luciferase activity of control (DMSO), different concentrations of GUDCA or TUDCA without or with FXR agonist CDCA treatment. n = 3 replicates/treatment. P value was determined by one-way ANOVA with Tukey’s correction (F7,16 = 61.69). c, SHP and FGF19 mRNA expression in differentiated Caco-2 cells after treatment with different concentrations of GUDCA or TUDCA with CDCA. n = 3 replicates/treatment. P value was determined by one-way ANOVA with Tukey’s correction (SHP: F7,16 = 4.871, FGF19: F7,16 = 15.41). d, Relative expression of the target gene mRNAs of intestinal FXR in mice treated with vehicle, TCA or TCA and CDCA, GUDCA or TUDCA. n = 5 mice/group. P value was determined by one-way ANOVA with Tukey’s correction (Shp: F4,20 = 38.47, Fgf15: F4,20 = 25.02). e, Relative expression of intestinal Fxr mRNA and its target gene mRNAs in mice treated with metformin on a HFD for 1 week. n = 5 mice/group. P value was determined by two-tailed Student’s t-test (Shp: t8 = 2.779, Fgf15: t8 = 3.383). f, The relative expression of Cyp7a1, Cyp7b1, Cyp8b1 and Cyp27a1 mRNAs in the livers of metformin-treated mice after 1-week HFD feeding. n = 5 mice/group. P value was determined by two-tailed Student’s t-test (Cyp7a1: t8 = 2.805). g, Relative abundance of intestinal Fxr mRNA and its target gene mRNAs in metformin-treated Ampka1fl/fl and Ampka1ΔIE mice on a HFD for 1 week. n = 5 mice/group. P value was determined by one-way ANOVA with Tukey’s correction (Shp: F3,16 = 8.046, Fgf15: F3,16 = 11.83). h, Relative expression of intestinal Fxr mRNA and its target gene mRNAs in antibiotics-treated, microbiota-depleted mice treated with metformin on a HFD for 1 week. n = 5 mice/group. P value was determined by one-way ANOVA with Tukey’s correction (Shp: F3,16 = 6.859, Fgf15: F3,16 = 8.782). i, In microbiota-depleted mice, metformin had no effect in inhibiting the activation of intestinal FXR on top of TCA. The relative expression of Fxr and its target genes in the ileum. n = 6 mice/group. All the data are presented as the mean ± s.e.m.

Article Snippet: After 24 h post-transfection, the cells were exposed to GUDCA (50–200 μM, Sigma-Aldrich, Cat# 06863), or TUDCA (50–200 μM, Sigma-Aldrich, Cat# T0266) in the presence of CDCA (20 μM, Targetmol, Cat# T0847) for 16 h. Luciferase assays were performed using the dual-luciferase assay system (Promega).

Techniques: Luciferase, Activity Assay, Expressing, Two Tailed Test, Activation Assay

Journal: Biomolecules

Article Title: Production of New Microbially Conjugated Bile Acids by Human Gut Microbiota

doi: 10.3390/biom12050687

Figure Lengend Snippet: BAs identified and confirmed of classical conversion trend of secondary BAs.

Article Snippet: Authentic standards of 3α,7α-Dihydroxy-5β-cholan-24-oic Acid (chenodeoxycholic acid) and 3α-Hydroxy-11-oxo-5β-cholan-24-oic Acid (3-oxo-chenodeoxycholic acid) were purchased from Avanti Polar Lipids (Alabaster, AL, USA).

Techniques:

Journal: Biomolecules

Article Title: Production of New Microbially Conjugated Bile Acids by Human Gut Microbiota

doi: 10.3390/biom12050687

Figure Lengend Snippet: MCBAs of valine identified. ( a ) valoisolithocholic acid; ( b ) valoiso7β-Hydroxy-5β-cholan-24-oic acid; ( c ) valolithocholic acid; ( d ) valoisolithocholate ester; ( e ) valoiso7β-Hydroxy-5β-cholate ester.

Article Snippet: Authentic standards of 3α,7α-Dihydroxy-5β-cholan-24-oic Acid (chenodeoxycholic acid) and 3α-Hydroxy-11-oxo-5β-cholan-24-oic Acid (3-oxo-chenodeoxycholic acid) were purchased from Avanti Polar Lipids (Alabaster, AL, USA).

Techniques:

Journal: Nature Communications

Article Title: Farnesoid X receptor activation by bile acids suppresses lipid peroxidation and ferroptosis

doi: 10.1038/s41467-023-42702-8

Figure Lengend Snippet: a Overexpression of FXR in HT-1080. Levels of mRNA were normalized to GAPDH mRNA. FXR levels are reverted to control levels upon treatment with 50 µM FXR inhibitor Guggulsterone for 24 h. Data are mean ± SD of n = 3 biological replicates; one-way ANOVA with Dunnett’s test. b FXR overexpression leads to elevated mRNA levels of anti-ferroptotic genes GPX4, FSP1, PPARα, ACSL3 and SCD1. Expression levels are normalized to mRNA levels of GAPDH. Upregulated anti-ferroptotic target genes in HT-1080 overexpressing FXR can be reverted to control levels by treatment with 50 µM FXR inhibitor Guggulsterone for 24 h. RP2 was used as a housekeeper gene for normalization. Data are mean ± SD of n = 3 biological replicates; one-way ANOVA with Dunnett’s test. c , d Ferroptosis inhibition by FXR activation is reverted when FSP1 or PPARα are inhibited. Ferroptosis was induced by 200 nM RSL3 for 18 h, 12 µM Turofexorate was used to inhibit ferroptosis. FSP1 was inhibited by iFSP1, PPARα was inhibited by GW6471 (PPARαi). Data are normalized to untreated control and plotted as mean ± SD of n = 3 biological replicates; one-way ANOVA with Tukey’s test. e Endogenous FXR activation by bile acids inhibits ferroptosis in HepG2 cells. Cells were treated with 65 nM RSL3, 20 µM Chenodeoxycholic acid (CDC) and 2 µM Obeticholic acid (OC) for 5 h. Data are mean ± SD of n = 6 biological replicates; one-way ANOVA with Dunnett’s test. f Levels of FXR and anti-ferroptotic genes GPX4, FSP1, PPARα, ACSL3 and SCD1 are elevated after bile acid treatment of HepG2. Cells were treated with 20 µM Chenodeoxycholic acid (CDC) or 2 µM Obeticholic acid (OC) for 8 h. Expression levels are normalized to mRNA levels of GAPDH. Data are mean ± SD of n = 4 biological replicates; one-way ANOVA with Dunnett’s test; D = DMSO.

Article Snippet: For experiments with endogenous FXR agonists: Chenodeoxycholic Acid (endogenous bile acid, FXR agonist, LKT Labs) and Obeticholic Acid (cholic acid derivative, FXR agonist, BioVision).

Techniques: Over Expression, Control, Expressing, Inhibition, Activation Assay