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Genome-wide maps of RNA Polymerase II and p65 localization in HUVECs stimulated with TNF alpha

ABSTRACT: TNF alpha is one of the inflammatory mediator and induce genes mainly by transcriptional factor, p65, in endothelial cells. This time, we performed a time course study to detect the change of localization of p65 and Pol II.

ORGANISM(S): Homo sapiens

PROVIDER: GSE34500 | GEO | 2013/01/01

SECONDARY ACCESSION(S): PRJNA151313

REPOSITORIES: GEO

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Genome-wide maps of RNA Polymerase II and p65 localization in HUVECs stimulated with TNF alpha

Project description:TNF alpha is one of the inflammatory mediator and induce genes mainly by transcriptional factor, p65, in endothelial cells. This time, we performed a time course study to detect the change of localization of p65 and Pol II. To identify p65 and Pol II binding sites, we used chromatin immunoprecipitation with deep sequencing (ChIP-seq) of HUVECs treated with or without TNF alpha for 30 mins. Cells were starved before stimulation longer than 16 hours. HUVECs were used within the first 6 passages. For crosslinking, 10 mM of EGS in 50% glacial acetic acid was used for 45 min, followed by 20 min of 1% paraformaldehyde treatmet was used.

2013-01-01 | E-GEOD-34500 | biostudies-arrayexpress

p65

Project description:p65

| PRJNA811604 | ENA

ENCODE ChIP-chip for Rel A (p65) using polyclonal antibody (N-) on TNF-alpha-stimulated HeLaS3 Cells [unpublished]

Project description:Rel A (p65) ChIP-chip using polyclonal antibody (N-terminus) performed on TNF-alpha-stimulated HeLaS3 cells for Nimblegen ENCODE arrays which comprise 50mer oligonucleotides spaces every 38bps (overlapping by 12nts). Goal was to identify p65-binding regions in the inflammatory and immune response of human cells. Keywords: ChIP-chip

2007-02-14 | GSE6900 | GEO

ENCODE ChIP-chip for Rel A (p65) using polyclonal antibody (C-20) on TNF-alpha-stimulated HeLaS3 Cells [unpublished]

Project description:Rel A (p65) ChIP-chip using polyclonal antibody (C-20) performed on TNF-alpha-stimulated HeLaS3 cells for Nimblegen ENCODE arrays which comprise 50mer oligonucleotides spaces every 38bps (overlapping by 12nts). Goal was to identify p65-binding regions in the inflammatory and immune response of human cells. Keywords: ChIP-chip

2007-02-14 | GSE6899 | GEO

Genome-wide maps of H3K27ac localization in HUVECs

2013-01-01 | GSE38555 | GEO

Genome-wide maps of H3K27ac localization in HUVECs

Project description:H3K27Ac is one of the expressed enhancer markers in endothelial cells, but its genomic localization is unknown. This time, we established a new antibody for H3K27ac, and performed ChIP-seq to identify H3K27ac binding site in whole genome manner under hypoxia. We used chromatin immunoprecipitation with deep sequencing (ChIP-seq) of HUVECs treated with or without hypoxia (1%O2) for 24hours, then H3K27ac binding regions were identified. Normoxia was used as a control condition. HUVECs were used within the first 6 passages.

2013-01-01 | E-GEOD-38555 | biostudies-arrayexpress

Genome-wide maps of MEF2C and H3K27ac localization in HUVECs

2013-01-01 | GSE32644 | GEO

Genome-wide maps of MEF2C and H3K27ac localization in HUVECs

Project description:MEF2C is one of the substantially expressed transcriptional factors in endothelial cells, but its genomic localization is unknown. This time, we established a new antibody for MEF2C, and performed ChIP-seq to identify MEF2C binding site in whole genome manner. H3K27Ac binding sites were also detected in the same way. We used chromatin immunoprecipitation with deep sequencing (ChIP-seq) of HUVECs treated with or without pitavastatin for 4hours, we identified MEF2C and H3K27Ac binding regions.HUVECs were used within the first 6 passages. For MEF2C studies, HUVECs were cultivated in medium EGM2MV containing pitavastatin at a concentration of 1 ?M, and same concentration of DMSO was used as a control sample. For H3K27ac, HUVECs were starved for 16 hours and harvested without statin treatment.

2013-01-01 | E-GEOD-32644 | biostudies-arrayexpress

p65-HaloTag copurifying proteins in human 293T cells

Project description:In effort to develop methodology for targeted top down mass spectrometry of NF kappa B p65 from human cells, we evaluated the utility of HaloTag for purification and analysis of recombinant protein. During our study, two datasets of bottom up LC-MS/MS were generated: one from in-gel digestion of the predominant band following p65-HaloTag purification, another from in-solution digestion of all the proteins present in a p65-HaloTag purification. p65-HaloTag copurifying proteins identified in our datasets include the known interactors c-Rel, NF-kappaB p105, NF-kappaB p100, and NF-kappaB inhibitor beta. Over 100 proteins were identified by at least two peptides using a Mascot ion cut-off score of 30.

2018-10-19 | PXD001932 | Pride

Analysis of glucocorticoid receptor (NR3C1), NFkb (p65 subunit), and RNA polymerase 2 (RNAP2) binding in Beas-2B airway epithelial cells

Project description:The objective of this study was to determine binding patterns for GR, 65 and RNAP2 in Beas-2B airway epithelial cells after treatment with dexamethasone (100 nm), TNF (20 ng/ml) or both for one hour. This study utilized duplicate samples for each treatment condition and immunoprecipitation except for p65 immunoprcipitation of TNF treated samples, which was analyzed as a single sample. Input from vehicle treated cells was used a control. Experimental comparisons were made between the following samples/ treatment conditions. Duplicate samples of Beas-2B cells treated with ethanol (vehicle) were used for ChIP of GR, p65, and RNAP2 (2 samples each antibody). Duplicate samples of Beas-2B cells treated with dexamethasone were used for ChIP of GR and RNAP2 (2 samples each antibody). Duplicate samples of Beas-2B cells treated with TNF were used for ChIP of p65 and RNAP2 (1 sample p65, 2 samples RNAP2). Duplicate samples of Beas-2B cells treated with dexamethasone + TNF were used for ChIP of GR, p65 and RNAP2 (2 samples each antibody).

2016-04-01 | E-GEOD-79803 | biostudies-arrayexpress

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