Journal: Oncology reports

Article Title: Farnesoid X receptor inhibits proliferation of human colorectal cancer cells via the miR‑135A1/CCNG2 signaling pathway.

doi: 10.3892/or.2018.6636

Figure Lengend Snippet: Figure 4. miR‑135A1 regulates proliferation by inhibiting CCNG2 in vitro. (A) Analysis of CCNG2 mRNA expression in CRC cells transfected with CCNG2 siRNA. (B) Western blot analysis of CCNG2 protein expression in CRC cells treated with CCNG2 siRNA. (C) Colony formation assay of CRC cells treated with miR‑135A1 inhibitor or CCNG2 siRNA.

Article Snippet: CCNG2 siRNA (50 nM), FXR siRNA (50 nM) and non‐specific scrambled siRNA (NC‐siRNA; 50 nM) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and transfected into SW620 or HCT116 cells for 48 h using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), as previously described (18).

Techniques: In Vitro, Expressing, Transfection, Western Blot, Colony Assay

Journal: Oncology reports

Article Title: Farnesoid X receptor inhibits proliferation of human colorectal cancer cells via the miR‑135A1/CCNG2 signaling pathway.

doi: 10.3892/or.2018.6636

Figure Lengend Snippet: Figure 4. Continued. (D) Colony formation of CRC cells treated with miR‑135A1 mimics or CCNG2 siRNA. (E) Cell cycle analysis was conducted following treatment of CRC cells with miR‑135A1 inhibitor or CCNG2 siRNA by flow cytometry. *P<0.05, ***P<0.001 vs. normal control; ###P<0.001 vs. miR‑135A1 inhibitor treatment group. miR, microRNA; CCNG2, cyclin G2; CRC, colorectal carcinoma; siRNA, small interfering RNA.

Article Snippet: CCNG2 siRNA (50 nM), FXR siRNA (50 nM) and non‐specific scrambled siRNA (NC‐siRNA; 50 nM) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and transfected into SW620 or HCT116 cells for 48 h using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), as previously described (18).

Techniques: Cell Cycle Assay, Flow Cytometry, Control, Small Interfering RNA

Journal: Oncology reports

Article Title: Farnesoid X receptor inhibits proliferation of human colorectal cancer cells via the miR‑135A1/CCNG2 signaling pathway.

doi: 10.3892/or.2018.6636

Figure Lengend Snippet: Figure 5. miR‑135A1 is involved in FXR suppression of proliferation by inducing CCNG2 expression in CRC cells. (A) A GW4064 dose‑response study was conducted for 24 h, and mRNA expression of miR‑135A1 was analyzed. (B) Western blot analysis of FXR protein expression in CRC cells treated with FXR siRNA. (C) Analysis of FXR mRNA expression in CRC cells treated with FXR siRNA. (D) Analysis of miR‑135A1 expression following transfection of SW620 and HCT116 cells with FXR siRNA and treatment with or without GW4064 for 24 h. (E) CCNG2 expression was upregulated depending on the concentration of GW4064. (F) Evaluation of CCNG2 protein expression by western blotting subsequent to transfection with FXR siRNA and treatment with or without GW4064 for 24 h.

Article Snippet: CCNG2 siRNA (50 nM), FXR siRNA (50 nM) and non‐specific scrambled siRNA (NC‐siRNA; 50 nM) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and transfected into SW620 or HCT116 cells for 48 h using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), as previously described (18).

Techniques: Expressing, Western Blot, Transfection, Concentration Assay

Journal: Oncology reports

Article Title: Farnesoid X receptor inhibits proliferation of human colorectal cancer cells via the miR‑135A1/CCNG2 signaling pathway.

doi: 10.3892/or.2018.6636

Figure Lengend Snippet: Figure 5. Continued. (F) Evaluation of CCNG2 protein expression by western blotting subsequent to transfection with FXR siRNA and treatment with or without GW4064 for 24 h. (G) Cells were transfected with miR‑135A1 inhibitor with or without GW4064 treatment for 24 h, and CCNG2 expression was analyzed by western blotting. (H) Colony formation ability was analyzed following transfection with miR‑135A1 inhibitor, with or without GW4064 treatment for 24 h. (I) CRC‑cell viability was analyzed by cell counting kit‑8 analysis in response to miR‑135A1 inhibitor transfection or GW4064 treatment. *P<0.05, ***P<0.01 vs. normal control; ###P<0.01 vs. GW4064 treatment group. miR, microRNA; FXR, farnesoid X receptor; CRC, colorectal carcinoma; siRNA, small interfering RNA; CCNG2, cyclin G2; NC, normal control; in, inhibitor.

Article Snippet: CCNG2 siRNA (50 nM), FXR siRNA (50 nM) and non‐specific scrambled siRNA (NC‐siRNA; 50 nM) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and transfected into SW620 or HCT116 cells for 48 h using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), as previously described (18).

Techniques: Expressing, Western Blot, Transfection, CCK-8 Assay, Control, Small Interfering RNA

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: Fig. 2 | Activation of FXR suppresses ferroptosis in cooperation with RXR. a Treatment of HT-1080 with 12 µM Turofexorate or Fexaramine for 7 h increases FXR expression. Levels of mRNA were normalized to GAPDH expression. Data are mean ± SD of n = 3 biological replicates. Western Blot shown is one representative experiment from n = 3. One-way ANOVA with Dunnett’s test; D = DMSO b Knockdown of FXR sensitizes cells to ferroptosis. HT-1080 were transfected with 40 nM esiRNA against FXR for 48 h and treated with 1 µM IKE for 18 h. EsiGFP- transfected cells served as a control group. Data are mean ± SD of n = 3 biological replicates; one-way ANOVA with Tukey’s test. c, d Turofexorate and Fexaramine are

Article Snippet: After 24 h of growth, cells were transfected with a vector expressing GFP-tagged human FXR (NM_001206979, OriGene) by usingX-tremeGENEHPDNATransfectionReagent (Sigma-Aldrich).

Techniques: Activation Assay, Expressing, Western Blot, Knockdown, Transfection, esiRNA, Control

Journal: Theranostics

Article Title: Gut microbiota depletion and FXR inhibition exacerbates zonal hepatotoxicity of sunitinib

doi: 10.7150/thno.99926

Figure Lengend Snippet: FXR-autophagy pathway participated in sunitinib-induced hepatotoxicity. ( A ) H&E staining of mouse liver. ( B ) Representative image of liver. ( C-G ) Serum AST ( C ), ALT ( D ), ALP ( E ), TBA ( F ), and gall bladder weight ( G ) in FXR knockout mice. ( H-L ) Body weight ( H ), H&E staining of mouse liver ( I ), AST ( J ), ALT ( K ), and TBA ( L ) in Fxr fl/fl , Fxr ΔL , and Fxr ΔIE mice. ( M ) Correlation analysis between FXR and autophagy gene expression through RNA-Seq analysis in wide type and FXR knockout mice. ( N ) Correlation analysis between FXR and autophagy gene expression in mouse liver. ( O ) Autophagy level using cell autophagy detection assay kit after 3-MA and NH 4 Cl treatment. ( P ) Cell viability and AST level after 3-MA treatment in mouse primary hepatocytes. ( Q ) Cell viability and AST level after NH 4 Cl treatment in mouse primary hepatocytes. ( R ) FXR (FXR, BSEP, SHP, and OSTβ) and autophagy (LC3BI/II and P62) protein expression in mouse liver after sunitinib treatment. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following antibodies were used: platelet endothelial cell adhesion molecule 1 (CD31, ab7388, Abcam, UK), CD31 (ab24590, Abcam, UK), cytokeratin 19 (CK19, ab254186, Abcam, UK), microtubule- associated protein light chain 3B (LC3B, ab192890, Abcam, UK), zonula occludens-1 (ZO-1, ab96587, Abcam, UK), FXR (NB400-153, Novus), FXR (sc-25309, Santa Cruz Biotechnology, USA), BSEP (PB0379, BOSTER), sequestonsome1 (SQSTM1/P62, ab109012, Abcam, UK), Caspase3 (ab184787, Abcam, UK).

Techniques: Staining, Knock-Out, Gene Expression, RNA Sequencing, Detection Assay, Expressing

Journal: Nature

Article Title: FXR inhibition may protect from SARS-CoV-2 infection by reducing ACE2.

doi: 10.1038/s41586-022-05594-0

Figure Lengend Snippet: Fig. 1 | FXR modulates ACE2 expression and SARS-CoV-2 infection. a, ChIP–qPCR on cholangiocyte organoids, showing that the FXR agonist CDCA promotes the binding of FXR on the ACE2 promoter, and that this is reduced by FXR inhibitors (UDCA and ZGG). OSTα as positive control; ACE2 promoter adjoining region as negative control; n = 4 independent experiments; one-way ANOVA adjusted for multiple comparisons; bars,s.d. b, Schematic representation of the suggested mechanism for FXR-mediated control of ACE2 expression and SARS-CoV-2 infection relative to e,f. c,d, qPCR (c) and immunofluorescence (d) showing the levels of ACE2 after modulation of FXR activity in primary airway, biliary and intestinal organoids. Housekeeping gene, HMBS (also known as PBGD); n = 4 independent experiments; one-way ANOVA; centre line,

Article Snippet: These gene reporter constructs were co-transfected with a commercially available FXR expression plasmid (OriGene, SC329876) into HEK293 cells using TransIT-293 Transfection Reagent (MirusBio).

Techniques: Expressing, Infection, ChIP-qPCR, Binding Assay, Positive Control, Negative Control, Control, Immunofluorescence, Activity Assay

Journal: Nature

Article Title: FXR inhibition may protect from SARS-CoV-2 infection by reducing ACE2.

doi: 10.1038/s41586-022-05594-0

Figure Lengend Snippet: Fig. 2 | Inhibition of FXR reduces ACE2 expression and SARS-CoV-2 infection in vivo. a, Schematic of the experiment performed in Syrian golden hamsters. Sentinel hamsters were not directly inoculated with virus. SARS-CoV-2 infection in sentinel hamsters was achieved through transmission from directly inoculated hamsters after co-housing. b, qPCR showing that treatment with UDCA reduces the levels of ACE2 in hamster nasal turbinates and lungs. Housekeeping gene, Gapdh; n = 5 vehicle (no UDCA) group versus n = 3 UDCA group; unpaired two-tailed t-test; centre line, median; box, interquartile range; whiskers, range; bars, s.d. c, Immunofluorescence images showing the levels of ACE2 in nasal and respiratory epithelium of hamsters receiving UDCA versus vehicle. n = 3 hamsters per group. Scale bars, 100 μm. d, qPCR showing the levels of SARS-CoV-2 RNA in swabs, nasal turbinates and lungs of directly inoculated hamsters and sentinel hamsters treated with UDCA or vehicle and co-housed with infected hamsters. Samples were collected after four days of co-housing. SARS-CoV-2 nucleocapsid RNA quantification relative to 18s rRNA. n = 3

Article Snippet: These gene reporter constructs were co-transfected with a commercially available FXR expression plasmid (OriGene, SC329876) into HEK293 cells using TransIT-293 Transfection Reagent (MirusBio).

Techniques: Inhibition, Expressing, Infection, In Vivo, Virus, Transmission Assay, Two Tailed Test, Immunofluorescence

Journal: Liver international : official journal of the International Association for the Study of the Liver

Article Title: Disentangling Organ-Specific Roles of Farnesoid X Receptor in Bile Acid and Glucolipid Metabolism.

doi: 10.1111/liv.70027

Figure Lengend Snippet: FIGURE 5 | Intestinal FXR improved hypertriglyceridemia by limiting lipid absorption. (A) Plasma TG concentrations and AUC in HFD-fed mice after olive oil gavage, n = 6–8. (B) Total daily faecal TG excretion and (C) total BAs excretion in 24 h, n = 5–7 per group. (D) Venn diagram of iden- tifying 90 DEGs regulated by intestinal FXR across groups. (E) KEGG pathway analysis of the 90 DEGs from (D). (F) Heatmap of DEG expression in the pathways highlighted in (B). (G) Relative mRNA expression of ileum Mttp and Fabp1 of different genotype mice fed on an HFD (n = 5). (H) Western blot analysis of MTTP and FXR from Caco2 cell lysates following adenovirus-mediated FXR overexpression or lentivirus-mediated FXR- knockdown. (I) TG transport experiment in differentiated Caco2 cells cultured on a transwell plate for 21 days; Left panel, schematic diagram; Right panel, TG content of the culture medium in the lower chambers of transwell plates were measured in Caco2 cells treated with Lomi or DMSO. Data were presented as mean ± SEM. *p < 0.05, versus WT; #p < 0.05, versus FxrRAΔNull; one-way ANOVA. Ad, adenovirus; BAs, bile acids; DEGs, differ- entially expressed genes; DMSO, dimethyl sulfoxide; KEGG, Kyoto Encyclopedia of Genes and Genomes; Lomi, lomitapide; Lv, lentivirus; OA, oleic acid; OLTT, oral lipid tolerance test; TG, triglyceride.

Article Snippet: The antibodies used were as follows: anti- FXR (PP- A9033A- 00, R&D Systems, USA), anti- MTTP (ab63467, Abcam, UK), anti- HSP90 (Santa Cruz Biotechnology, USA).

Techniques: Clinical Proteomics, Expressing, Western Blot, Over Expression, Knockdown, Cell Culture