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Image Search Results
Journal: BMC Gastroenterology
Article Title: Mapping of HNF4α target genes in intestinal epithelial cells
doi: 10.1186/1471-230X-9-68
Figure Lengend Snippet: Analysis of the CDX2 promoter . A) The promoter region cloned in pGL4.10. The coordinates are relative to +1, the transcriptional start site. The arrows indicate primers used in the real-time qPCR. The possible HNF4α binding site and the mutated sequence is shown below. B) HNF4α ChIP-chip and AcHis3 ChIP-chip results for probes spanning the CDX2 promoter. Enrichments are shown as Log2 ratios between immunoprecipitated and input DNA. N = 3. C) Real-time qPCR with CDX2 and IgG intron primers using HNF4α ChIP, HA ChIP and input DNA. Enrichments are presented as percent of total input. N = 4. The CDX2 promoter enrichment is statistical significant (p-values < 0.05, Student T test). D) Gel shift analysis of the HNF4α site in the CDX2 promoter using Caco-2 nuclear extract (lane 1). Competition by unlabelled wt (lane 2), mut-HNF4α oligonucleotides (lane 3) and an unrelated oligonucleotide (lane 4) demonstrating a specific binding. One of the two protein/DNA bands can be supershifted with HNF4α antibody (lane 5). E) Promoter analysis of CDX2 promoter in Caco-2 (grey bars) and COS7 (white bars) cells with and without co-transfection of HNF4α expression plasmid. The binding site for HNF4α was mutated in order to analyze the functional importance of the HNF4α site (pGL4-CDX2 mutHNF4). The luciferase activity was corrected for transfection efficiency and normalized to the expression of pGL4-CDX2, N = 4.
Article Snippet: The amplified
Techniques: Clone Assay, Binding Assay, Sequencing, ChIP-chip, Immunoprecipitation, Gel Shift, Cotransfection, Expressing, Plasmid Preparation, Functional Assay, Luciferase, Activity Assay, Transfection
Journal: BMC Gastroenterology
Article Title: Mapping of HNF4α target genes in intestinal epithelial cells
doi: 10.1186/1471-230X-9-68
Figure Lengend Snippet: Analysis of the trehalase ( TREH ) promoter . A) Map of the TREH promoter region. PCR primers, the HNF4α binding site, and the mutations are shown. B) HNF4α ChIP-chip and AcHis3 ChIP-chip results for probes spanning the TREH promoter. N = 3. C) Real-time qPCR analysis of HNF4α ChIP, HA ChIP and input DNA using TREH and IgG intron primers. N = 3. The TREH promoter enrichment is statistical significant (p-values < 0.05, Student T test). D) Gel shift analysis of the HNF4α site in the TREH promoter. E) Promoter analysis of the TREH promoter in Caco-2 (grey bars) and COS7 (white bars) cells with and without co-transfection of HNF4α expression plasmid and a mutation in the HNF4α site (pGL4-TREH mutHNF4). The luciferase activity was corrected for transfection efficiency and normalized to the expression of pGL4-TREH, N = 4.
Article Snippet: The amplified
Techniques: Binding Assay, ChIP-chip, Gel Shift, Cotransfection, Expressing, Plasmid Preparation, Mutagenesis, Luciferase, Activity Assay, Transfection
Journal: BMC Gastroenterology
Article Title: Mapping of HNF4α target genes in intestinal epithelial cells
doi: 10.1186/1471-230X-9-68
Figure Lengend Snippet: Analysis of the cingulin ( CGN ) promoter . A) Map of the CGN promoter region. PCR primers, the HNF4α binding site, and the mutations are shown. B) HNF4α ChIP-chip and AcHis3 ChIP-chip results for probes spanning the CGN promoter. N = 3. C) Real-time qPCR analysis of HNF4α ChIP, HA ChIP and input DNA using CGN and IgG intron primers. N = 3. The CGN promoter enrichment is statistical significant (p-values < 0.05, Student T test). D) Gel shift analysis of the HNF4α site in the CGN promoter. E) Promoter analysis of the CGN promoter in Caco-2 (grey bars) and COS7 (white bars) cells with and without co-transfection of HNF4α expression plasmid and a mutation in the HNF4α site (pGL4-CGN mutHNF4). The luciferase activity was corrected for transfection efficiency and normalized to the expression of pGL4-CGN, N = 4.
Article Snippet: The amplified
Techniques: Binding Assay, ChIP-chip, Gel Shift, Cotransfection, Expressing, Plasmid Preparation, Mutagenesis, Luciferase, Activity Assay, Transfection
Journal: BMC Gastroenterology
Article Title: Mapping of HNF4α target genes in intestinal epithelial cells
doi: 10.1186/1471-230X-9-68
Figure Lengend Snippet: HNF4α ChIP on mouse small intestinal epithelium . Chromatin immunoprecipitation analysis of HNF4α binding to HNF1α ( Tcf1 ), phosphoenolpyruvate carboxykinase 1 ( Pck1 ), apolipoprotein C3 ( Apoc3 ), Cdx-2 ( Cdx2 ), trehalase ( Treh ), and cingulin ( Cgn ) promoters in mouse small intestinal epithelium. An intron in the IgG gene served as a negative control (IgG). The analyzed promoter regions are all conserved between human and mouse. Enrichments are represented as percent of the total amount of genomic input DNA in ChIP. Significant enrichments are indicated (p-values < 0.001 is shown by ***, p-values < 0.05 is shown by *).
Article Snippet: The amplified
Techniques: Chromatin Immunoprecipitation, Binding Assay, Negative Control