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WGBS DNA methylation alignments of differentiated HL60/s4 cells

ABSTRACT: Whole-genome fingerprint of the DNA methylome during chemically induced differentiation of the human AML cell line HL-60/S4

PROVIDER: PRJEB27665 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR5729412.fastq.gz	Fastqsanger.gz
	ERR5729412_1.fastq.gz	Fastqsanger.gz
	ERR5729412_2.fastq.gz	Fastqsanger.gz
	ERR5729413.fastq.gz	Fastqsanger.gz
	ERR5729413_1.fastq.gz	Fastqsanger.gz

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Project description:Ubiquitin specifc protease (USP) 2 modulates various cellular responces including carcinogenesis, spermatogenesis, and glucose metabolism. So far, we established USP2 knockdown HL-60 cells to explore roles of USP2 in macrophages. In this experiment, we investigated whether USP2 knockdown HL-60 cells more prominently modulate function of adipocytes compared with control cells. USP2 knock down(KD) HL-60 cells and mock vector-transfected mock control cells were differentiated into macrophage-like cells in the presence of 30nM of phorbol 12-myristate 13-acetate for 5 days. Subseqently, supernatants of the USP2 KD cells and control cells were added to differentiated 3T3-L1 cells for 3 and 6 hrs. Total RNA was isolated from the supernatant-treated 3T3-L1 cells, and subjected to microarray analysis. Duplicate samples were prepared for each cell and time point.

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Project description:Cell differentiation is associated with changes in chromatin organization and gene expression. In this study, we examine chromatin structure following differentiation of the human myeloid leukemia cell line (HL-60/S4) into granulocytes with retinoic acid (RA) or into macrophage with phorbol ester (TPA). We performed ChIP-seq of histone H3 and its modifications, analyzing changes in nucleosome occupancy, nucleosome repeat length, eu-/heterochromatin redistribution and properties of epichromatin (surface chromatin adjacent to the nuclear envelope). Nucleosome positions changed genome-wide, exhibiting a specific class of alterations involving nucleosome loss in extended (~1kb) regions, pronounced in enhancers and promoters. Genes that lost nucleosomes at their promoters showed a tendency to be upregulated. On the other hand, nucleosome gain was not systematic at functional elements and did not show simple effects on transcript levels. The average genome-wide nucleosome repeat length (NRL) did not change significantly with differentiation. However, we detected an approximate 10 bp NRL decrease around the hematopoietic transcription factor (TF) PU.1 and the architectural protein CTCF, suggesting an effect on NRL proximal to TF binding sites. Nucleosome occupancy changed in regions associated with H3K4me3 in differentiated cells, compared to untreated HL-60/S4 cells. Epichromatin regions, largely unaffected by cell differentiation, revealed an increased GC content and high nucleosome density compared to surrounding chromatin. Epichromatin showed depletion of major (permissive and repressive) histone modifications and revealed enrichment with PML body-associated genes. In general, chromatin changes during HL-60/S4 differentiation appeared to be more localized, compared to changes among diverse cell types studied elsewhere, suggesting that the cell lineage differentiation of HL-60/S4 cells involves less drastic nucleosome repositioning.

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