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chip-chromatin-immunoprecipitation

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Journal Articles
Journal: Diabetes
Diabetes 2008;57(12):3189–3198
Published: 01 December 2008
...— We used chromatin immunoprecipitation (ChIP) linked to microarray (ChIP-chip) approach to compare genome-wide histone H3 lysine 9 dimethylation (H3K9me2) patterns in blood lymphocytes and monocytes from type 1 diabetic patients versus healthy control subjects. Bioinformatics evaluation of methylated...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2010;59(1):153–160
Published: 15 October 2009
... hybridization signal in the ChIP-on-chip assay. The position of the ATG is shown in bold. B: MIN6 cells were incubated in 3 versus 30 mmol/l glucose for 16 h, as indicated before cross-linking and chromatin immunoprecipitation using our polyclonal anti-ChREBP antibody. The primer sequences used...
Includes: Supplementary data
Meeting Abstracts
Journal: Diabetes
Diabetes 2019;68(Supplement_1):1999-P
Published: 01 June 2019
... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) is a modified CRISPR/Cas9 methodology that identifies novel transcriptional regulators. EnChIP uses dCas9 (lacks endonuclease activity) and guide RNAs (gRNAs) targeting a specific promoter region. The DNA-bound dCas9 can...
Journal Articles
Journal: Diabetes
Diabetes 2012;61(5):1062–1071
Published: 13 April 2012
... gene in vitro using the reporter assay, the electromobility shift assay (EMSA), and the chromatin immunoprecipitation (ChIP) assay in HuH7 cells and also using the mouse liver in vivo. We found that the transcriptional activity of the PFK2/FBP2 gene was stimulated by insulin and inhibited by cAMP...
Journal Articles
Journal: Diabetes
Diabetes 2011;60(11):2883–2891
Published: 17 October 2011
... product was calculated. Data are expressed as ratio to the copy number of FoxO1 mRNA expression. The sequences of the specific primer pairs are described in Supplementary Table 1 . Chromatin immunoprecipitation (ChIP) assays were performed using the ChIP-IT Express kit (Active Motif) from...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2010;59(7):1674–1685
Published: 22 April 2010
... Rfx3 expression in Min6 β-cells. Quantitative chromatin immunoprecipitation (ChIP), ChIP sequencing, and bandshift experiments were used to identify Rfx3 target genes. RESULTS Reduced development of insulin-positive cells in Rfx3−/− mice was not due to deficiencies in endocrine...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2010;59(3):564–571
Published: 10 December 2009
... oxidation, lipolysis, adipokine secretion, and mRNA expression was determined in human adipocytes. The interaction between twist1 and specific promoters in human adipocytes was investigated by chromatin immunoprecipitation (ChIP) and reporter assays. RESULTS Twist1 was highly expressed in human WAT...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(10):2730–2736
Published: 01 October 2006
... DIABETES 2006 ChIP, chromatin immunoprecipitation assay KATP channel, ATP-sensitive K+ channel KRBB, Krebs-Ringer bicarbonate buffer TSS, transcriptional start site UCP2, uncoupling protein 2 Slides were blocked with Avidin D and Biotin blocking reagents (Vector...
Meeting Abstracts
Journal: Diabetes
Diabetes 2019;68(Supplement_1):475-P
Published: 01 June 2019
...-induced SR-BI promoter activity and AMPK phosphorylation. As a downstream of AMPK, a transcription factor FoxO1 was confirmed to directly bind to the SR-BI promoter region by Chromatin Immunoprecipitation (ChIP) assay and overexpression of FoxO1 enhanced SR-BI promoter activity. Moreover, exendin-4...
Meeting Abstracts
Journal: Diabetes
Diabetes 2019;68(Supplement_1):1891-P
Published: 01 June 2019
... hours. Chromatin immunoprecipitation(CHIP)-qPCR were used in assaying the enrichment for histone active marks H3K9ac in liver PNPLA3. Real-time PCR was used to measure the mRNA expression of PNPLA3, and SIRT1 which is a known deacetylase affected by nutritional status. Results: The results showed...
Meeting Abstracts
Journal: Diabetes
Diabetes 2018;67(Supplement_1):192-OR
Published: 01 July 2018
... experiments including immunostaining and chromatin immunoprecipitation with antibodies that recognize only ChREBPα or both isoforms. We found that ChREBPα is only transiently expressed in the nucleus. By contrast, ChREBPβ, once expressed at high enough levels to detect, is constitutively nuclear. Our data...
Images
<span class="search-highlight">ChIP</span> in SVF ex vivo. Whole epididymal fat pads were fixed with formaldehyde...
Published: 18 October 2013
FIG. 2. ChIP in SVF ex vivo. Whole epididymal fat pads were fixed with formaldehyde immediately after dissection. After SVF isolation, ChIP was performed with the use of anti-STAT3, CEBP-β, and anti-acetylated histone H3 (AcH3) (kbp 9–14) antibodies followed by gel electrophoresis of 50% of the associated DNA (A) and qRT-PCR of the remainder (B). Negative control is chromatin incubated with nonimmune rabbit IgG. Input indicates the level of specific target chromatin before immunoprecipitation and was used to normalize qRT-PCR values. A: Representative gel electrophoresis showing that HFD induces STAT3 recruitment to a ∼300-bp fragment of pfn 5′-UTR (−711 to −406), encompassing a single STAT3 response element (−505 to −513) in association with increased lysine acetylation of H3 (arrow). B: The quantification of the chromatin bound to candidate transcription factors within the pfn promoter. Compared with NC, HFD promotes a statistically significant STAT3 recruitment to the fragment of pfn 5′-UTR (−711 to −406) that encompasses a single STAT3 response element (−505 to −513) as well as increased lysine acetylation of H3 of the same fragment, suggesting transcriptional activation. CEBP-β did not associate with a more proximal fragment of pfn 5′-UTR encompassing two response elements (−302 to −315 and −182 to −192, respectively). For each ChIP, chromatin was pooled from three HFD or eight NC mice (n = 3–5 pools/group). *P < 0.0001. IP, immunoprecipitate. FIG. 2. ChIP in SVF ex vivo. Whole epididymal fat pads were fixed with formaldehyde immediately after dissection. After SVF isolation, ChIP was performed with the use of anti-STAT3, CEBP-β, and anti-acetylated histone H3 (AcH3) (kbp 9–14) antibodies followed by gel electrophoresis of 50% of the associated DNA (A) and qRT-PCR of the remainder (B). Negative control is chromatin incubated with nonimmune rabbit IgG. Input indicates the level of specific target chromatin before immunoprecipitation and was used to normalize qRT-PCR values. A: Representative gel electrophoresis showing that HFD induces STAT3 recruitment to a ∼300-bp fragment of pfn 5′-UTR (−711 to −406), encompassing a single STAT3 response element (−505 to −513) in association with increased lysine acetylation of H3 (arrow). B: The quantification of the chromatin bound to candidate transcription factors within the pfn promoter. Compared with NC, HFD promotes a statistically significant STAT3 recruitment to the fragment of pfn 5′-UTR (−711 to −406) that encompasses a single STAT3 response element (−505 to −513) as well as increased lysine acetylation of H3 of the same fragment, suggesting transcriptional activation. CEBP-β did not associate with a more proximal fragment of pfn 5′-UTR encompassing two response elements (−302 to −315 and −182 to −192, respectively). For each ChIP, chromatin was pooled from three HFD or eight NC mice (n = 3–5 pools/group). *P < 0.0001. IP, immunoprecipitate. More
Journal Articles
Journal: Diabetes
Diabetes 2013;62(3):875–886
Published: 14 February 2013
... and chromatin immunoprecipitation (ChIP) analyses demonstrated that many important Isl1-activated genes were coregulated by Ldb1, including MafA, Arx, insulin, and Glp1r. However, some genes (i.e., Hb9 and Glut2) only appeared to be impacted by Ldb1 during...
Includes: Supplementary data
Images
<span class="search-highlight">ChIP</span> assay. <em>A</em>: Effect of glucose or insulin on the SREBP-1...
Published: 01 June 2005
FIG. 6. ChIP assay. A: Effect of glucose or insulin on the SREBP-1 binding to putative GLUT2-SRE. Chromatin was precipitated using a SREBP-1 antibody from primary hepatocytes, and the GLUT2 promoter region was amplified by PCR. The amount of chromosomal DNA used in immunoprecipitation between groups was normalized by input chromatin (100th of chromosomal DNA used for immunoprecipitation). B: Effect of glucose concentrations on the SREBP-1 binding to putative GLUT2-SRE. Glucose concentrations are shown on the top. Detailed PCR conditions and methods of sample treatment are described in research design and methods. FIG. 6. ChIP assay. A: Effect of glucose or insulin on the SREBP-1 binding to putative GLUT2-SRE. Chromatin was precipitated using a SREBP-1 antibody from primary hepatocytes, and the GLUT2 promoter region was amplified by PCR. The amount of chromosomal DNA used in immunoprecipitation between groups was normalized by input chromatin (100th of chromosomal DNA used for immunoprecipitation). B: Effect of glucose concentrations on the SREBP-1 binding to putative GLUT2-SRE. Glucose concentrations are shown on the top. Detailed PCR conditions and methods of sample treatment are described in research design and methods. More
Journal Articles
Journal: Diabetes
Diabetes 2010;59(1):249–255
Published: 15 October 2009
...), and chromatin immunoprecipitation (ChIP) was performed ± GLO1 overexpression for NFκB p65 (RAGE promoter) and AP-1 (S100A8, S100A12, and HMGB1 promoters). Finally, endothelial cells from nondiabetic mice, STZ diabetic mice, and STZ diabetic mice treated with the superoxide dismutase mimetic Mn(III)tetrakis(4...
Journal Articles
Journal: Diabetes
Diabetes 2008;57(4):1069–1077
Published: 01 April 2008
... 28 8 2007 DIABETES 2008 ChIP, chromatin immunoprecipitation DAG, diacylglycerol EMSA, electrophoretic mobility shift assay HNF4α, hepatic nuclear factor 4α HNF1pro, HNF1α-promoter IP3, inositol-triphosphate PKC, proteinkinase C PLC, phospholipase C PLCB1, β1 isoform...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2005;54(4):1064–1073
Published: 01 April 2005
... correspondence and reprint requests to Ming-Jer Tsai, PhD, Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. E-mail: mtsai@bcm.tmc.edu 7 1 2005 16 9 2004 DIABETES 2005 CHIP, chromatin immunoprecipitation assay NNAT...
Journal Articles
Journal: Diabetes
Diabetes 2003;52(2):403–408
Published: 01 February 2003
... cell enriched, BETA2 (also known as NeuroD1) ( 18 ), and ubiquitously distributed E47. Chromatin immunoprecipitation (ChIP) analysis also showed that BETA2 binds in vivo to the βGK region. Because islet ( 19 ), gut ( 19 ), and brain development ( 20 , 21 ) were all severely...
Images
<span class="search-highlight">ChIP</span> assay. <em>A</em>: The nucleotide sequences of the primers for...
Published: 13 April 2012
FIG. 7. ChIP assay. A: The nucleotide sequences of the primers for PCR used in the assay. Primers recommended by the manufacturers’ instruction of the ChIP-IT Express chromatin immunoprecipitation kit was used as a negative control (see research design and methods ). B: HuH7 cells were transfected with LXRα/RXRα expression plasmids, and the cells were homogenized with Dounce-type homogenizer and then treated with anti-LXRα antibody. The protein-DNA complex was separated with protein G–coated beads, treated with protease K, and then applied for PCR amplification (for details, see research design and methods ). FIG. 7. ChIP assay. A: The nucleotide sequences of the primers for PCR used in the assay. Primers recommended by the manufacturers’ instruction of the ChIP-IT Express chromatin immunoprecipitation kit was used as a negative control (see research design and methods). B: HuH7 cells were transfected with LXRα/RXRα expression plasmids, and the cells were homogenized with Dounce-type homogenizer and then treated with anti-LXRα antibody. The protein-DNA complex was separated with protein G–coated beads, treated with protease K, and then applied for PCR amplification (for details, see research design and methods). More
Journal Articles
Journal: Diabetes
Diabetes 2004;53(2):321–329
Published: 01 February 2004
..., Sunnyvale, CA). Chromatin immunoprecipitation (ChIP) assay was performed according to Orlando et al. ( 19 ). Briefly, formaldehyde cross-linked chromatin extracts were prepared from human myotubes and fragmented by sonication. First, 20 μg of chromatin extract were precleared with protein A Sepharose...