ABSTRACT: Fxr+/+ or Fxr-/- mice were gavaged with vehicle or GW4064 (30 mg/kg, twice a day) for 7 days. Livers were collected. Hepatic mRNA levels were determined using Sentrix MouseRef-8 v2 Expression BeadChip.
Project description:Farnesoid X receptor (FXR) is a ligand activated nuclear receptor belonging to the nuclear receptor superfamily. Bile acids (BAs) are the endogenous ligand for FXR. FXR is a master regulator of BA homestasis, including BA synthesis, metabolism, transport, and enterohepatic circulation of BAs. Besides, FXR is involved in regulating diverse physioligical function in both humans and mice. GW4064 is a synthetic FXR agonist which selectively activates FXR and induce the transcription of FXR target genes.
Project description:Farnesoid X receptor (FXR) is a ligand activated nuclear receptor belonging to the nuclear receptor superfamily. Bile acids (BAs) are the endogenous ligand for FXR. FXR is a master regulator of BA homestasis, including BA synthesis, metabolism, transport, and enterohepatic circulation of BAs. Besides, FXR is involved in regulating diverse physioligical function in both humans and mice. GW4064 is a synthetic FXR agonist which selectively activates FXR and induce the transcription of FXR target genes. In this study, we treated wild type C57BL/6J mice with GW4064 at 100mg per kg body weight or vehicle control. Mice were treated three times, first dose at 8 am, second dose at 6 pm, third dose at 8 am the second day. Mice were sacrificed 2 hours after the last dose, and liver tissues were harvested for analysis. Animal protocols and procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Kansas Medical Center. Total RNA from livers was prepared with TRIzol Reagent (Invitrogen, CA), and the whole transcription expression levels were determined using Mouse Gene 1.0 ST Array system manufactured by Affymetrix, Inc..
Project description:We have employed whole genome microarray expression profiling with the samplaes prepared by treating BMDM with GW4064, an fxr agonist. Some genes upregulated in GW4064 treated sample were used for qPCR to confirm the change of expression level.
Project description:To identify genes regulated by FXR in GLUTag cell line, we compared gene expression profiles in GLUTag cells treated for 24h by 0.1% DMSO and by 5M-5M GW4064
Project description:Global gene expression in primary cultured mouse kidney proximal tubule cells treated with either DMSO or 1uM GW4064 (an FXR agonist) was compared. Results provide insight into mechanisms underlying effects of FXR activation on gene expression in mouse kidney proximal tubule cells.
Project description:Global gene expression in the primary cultured mouse kidney proximal tubule cells treated either DMSO or 1uM GW4064 (a FXR agonist) was compared. Results provide insight into mechanisms underlying effects of FXR activation on gene expression in mouse kidney proximal tubule cells. Male C57/BJ mice aged 6 weeks were sacrificed under anesthesia and kidney proximal tubule cells were cultured until confluent. Cells were treated with either GW4064 (1uM) or equal amount of DMSO and incubated for 24 hours. 4 total RNA samples per group were analyzed and gene expression was compared between the groups.
Project description:We studied the effect of bile acid CDCA, FXR agonsit GW4064 and FGF19 on the expression levels of microRNA in cultured human primary hepatocytes
Project description:Mouse primary hepatocytes (MPH) prepared from C57Bl6 male adult mice were challenged with the FXR agonist GW4064 and used to prepare RNA which was then processed for analysis using MoGene-2_0-st Affymetrix microarrays according to standard procedures.
Project description:Expression profiling of whole body (WB) FXR knockout (KO) mice (FXR WB KO), liver-specific FXR KO mice (AFXR Cre+) and enterocyte specific FXR KO mice (VFXR Cre+) on a C57BL/6J genetic background Whole body (WB) FXR knockout (KO) mice (FXR WB KO), liver-specific FXR KO mice (AFXR Cre+) and enterocyte specific FXR KO mice (VFXR Cre+) on a C57BL/6J genetic background were bred and maintained in the laboratory animal research facility at the University of Kansas Medical Center in rooms under a 12-hour light-dark cycle. All experiments used 10-16 week-old male mice. FXR was activated by treatment with the FXR synthetic agonist, GW4064, at 150 mg/kg. GW4064 or vehicle was administered by oral gavage at 6 p.m., followed by a second administration at 8 a.m. the next morning. Two hours later, the liver was harvested.
Project description:Purpose: The purpose of the study was to investigate the differential expression pattern of genes in Rag2 KO mice spleen compared to its wild type counterpart. Methods: In this study, we have excised the spleen tissues from three Rag2 KO mice and three wild type mice, and then RNA samples were prepared with the spleen tissues for the analysis of transcriptomic profiles using the Illumina MouseRef-8 v2 Expression BeadChip platforms. The MouseRef-8 v2.0 BeadChip Kit content is derived from the national center for biotechnology information reference sequence (NCBI RefSeq) database (Build 36, Release 22). The content was supplemented with probes derived from the mouse exonic evidence based oligonucleotide (MEEBO) set, as well as standard protein-coding sequences described in the RIKEN FANTOM2 database. As well, the MouseRef-8 v2.0 BeadChip targets approximately 25,600 well-annotated RefSeq transcripts, over 19,100 unique genes, and enables the interrogation of 8 samples in parallel. The MouseRef-8 v2.0 BeadChip Kit uses the DirectHyb assay and is compatible with the iScan, HiScan, and Bead array reader systems. Result: The genes expression BeadChip array analysis showed that many genes have been expressed differentially between Rag2 KO and wild type mice spleen. These genes might be involved in the different biological and physiological processes including immune regulations. Conclusion: The differential genes expression profile will provide important insight about the immune deregulation in Rag2 KO mice.