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Drosophila melanogaster

ABSTRACT: Nuclear Pores provide a scaffold for induced enhancer promoter contacts

PROVIDER: PRJNA374851 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5259250.fastq.gz	Fastqsanger.gz
	SRR5259260.fastq.gz	Fastqsanger.gz
	SRR5259251.fastq.gz	Fastqsanger.gz
	SRR5259261.fastq.gz	Fastqsanger.gz
	SRR5259252.fastq.gz	Fastqsanger.gz

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Project description:Here we improved BiTS-ChIP (Bonn et al, Nature Protocols 7, 978-994 (2012)) to identify active enhancer and promoter elements genome wide in the 104 cardiomyocytes that constitute the Drosophila heart tube and represents only ~0.5% of the total cell content of the embryo. A transgenic Drosophila strain expressing nuclear GFP under the control of a cardiac specific enhancer (TinC*>GFP) was used for staged embryo collections at stages 13-14 (10-13h of development). After embryo fixation and dissociation, intact fixed nuclei were fluorescent labelling.Â Â Purification of this rare nuclear population was achieved by a two-step sorting procedure, yielding ~98% purity. Chromatin was extracted and used for immunoprecipitation and sequencing (ChIP-seq) to analyze chromatin modifications at promoters (H3K4me3 and H3K27ac) and enhancers (H3K27ac).Â Â Two independent biological replicates (from FACS sorting, chromatin preparations and ChIP-Seq) were performed for each mark and sequenced using Illumina HiSeq.

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