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RNA-seq of primary aortic human endothelial cells before and after TNFα stimulation

ABSTRACT: RNA-seq of primary aortic human endothelial cells before and after TNFα stimulation

PROVIDER: PRJEB32275 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR3289030_1.fastq.gz	Fastqsanger.gz
	ERR3289030_2.fastq.gz	Fastqsanger.gz
	ERR3289031_1.fastq.gz	Fastqsanger.gz
	ERR3289031_2.fastq.gz	Fastqsanger.gz
	ERR3289032_1.fastq.gz	Fastqsanger.gz

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RNA-seq of primary aortic human endothelial cells before and after TNFα stimulation

Project description:To characterize the transcriptome of primary vascular endothelial cells (ECs) during TNFα-response, we performed total RNA-seq on primary human aortic ECs (HAEC), before and after TNFα (45 min. 10 ng/mL).

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ATAC-seq of primary mammalian aortic endothelial cells before and after TNFα stimulation

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Project description:TNFα has an evolutionary conserved role in mediating inflammation via activation of the transcription factor NF-κB. The functions of individual NF-κB binding sites are not well understood. To identify conserved and functionally important NF-κB binding sites in mammals, we performed ChIP-seq to map the genome-wide binding of RELA and select histone modifications in primary vascular endothelial cells (ECs) isolated from the aortas of human (HAEC), mouse (MAEC) and cow (BAEC), before and after TNFα. The conserved RELA binding sites show strong epigenetic changes in response to TNFα and enrich near genes controlling vascular development and pro-inflammatory responses. Our method identifies novel modes of RELA-chromatin interactions that are conserved in mammals and shared between multiple cell-types. Particularly, genomic regions bound by RELA prior to stimulation are important responders during TNFα stimulation. We use CRISPR/Cas9 genome editing to validate the roles of the conserved RELA pre-bound sites near pro-inflammatory genes such as CCL2 and PLK2. Our evolutionary approach describes new aspects of mammalian NF-κB biology including its role within super-enhancers and relevance in inflammatory disorders.

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Project description:To identify conserved TNFα-induced changes in chromatin-accessibility in mammals, we performed ATAC-seq in primary vascular endothelial cells (ECs) isolated from the aortas of human (HAEC), mouse (MAEC) and cow (BAEC), before and after TNFα. We overlay our data with multi-species NF-κB binding data and identify multiple modes of NF-κB-chromatin interactions that are conserved during mammalian TNFα response. Our cross-species approach identifies conserved changes in chromatin-accessibility at NF-κB binding sites that are disease-relevant and essential during mammalian acute inflammation.

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Histone ChIP-seq of primary mouse aortic endothelial cells treated with TNFα against untreated controls

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