Project description:Heat shock rapidly induces expression of a small set of genes while globally repressing transcription, making it an attractive system for studying alterations in the chromatin landscape that accompany changes in gene regulation. We have characterized these changes using low-salt extraction of intact micrococcal nuclease (MNase)-treated Drosophila S2 cell nuclei to determine the active nucleosomal and subnucleosomal chromatin landscapes. The low-salt-soluble fraction corresponds to classical "active" chromatin and includes distinct size fractions of MNase-protected particles that can be precisely mapped by paired-end sequencing. After heat shock, the distribution of low-salt-soluble nucleosomes showed an overall reduction over gene bodies, consistent with down-regulation of transcription. No global changes were detected in the subnucleosomal landscape upstream of transcriptional start sites, however, we observed a genome-wide reduction of paused RNA Polymerase II from the active chromatin fraction. Furthermore, nucleosome turnover decreased within gene bodies in a pattern similar to that observed when transcription elongation was artificially inhibited. These observations suggest that reduced Pol II affinity and processivity is the dominant nuclear mechanism for genome-wide repression during heat shock. Our ability to precisely map both nucleosomal and subnucleosomal particles directly from classical active chromatin extracts to assay changes in the chromatin landscape provides a simple general strategy for epigenome characterization. High-throughput sequencing (Illumina HiSeq 2000) We have characterized changes to the active nucleosomal and subnucleosomal landscape during the heat shock response in Drosophila cells by genome-wide profiling of low-salt extracted micrococcal nuclease-treated nuclei, paused RNA Polymerase II and CATCH-IT nucleosome turnover.
Project description:To investigate the effect on chromatin accessibility with reduced ERBB2 signalling in oesophageal adenocarcinoma, we performed ATAC-seq in OE19 cells treated with either siNT or siERBB2.
Project description:To investigate the effects of gene expression of reduced ERBB2 signalling in oesophageal adenocarcinoma, we performed RNA-seq of OE19 cells treated with either siNT or siERBB2.
Project description:To investigate changes to gene expression associated with resistance to lapatinib, we performed ATAC-seq on OE19 cells treated with 500 nM lapatinib for 1, 7 and 35 days and vehicle control (DMSO) for 1 day.
Project description:The genome-wide binding sites of transcription factors provide insight into their regulatory function. We used chromatin immunoprecipitaition with high-throughput sequencing (ChIP-seq) to identify the genomic binding sites of HNF4A and GATA6 in OE19 cells.
Project description:To investigate changes to chromatin accessibility associated with resistance to lapatinib, we performed ATAC-seq on OE19 cells treated with 500 nM lapatinib for 1, 7 and 35 days and vehicle control (DMSO) for 1 day.
Project description:To determine genes regulated by the transcriptional co-activator PPARGC1A in OAC cells and whether PPARGC1A activity is altered by lapatinib we performed RNA-seq on OE19 cells treated with non-targeting siRNA or siRNA targeting PPARGC1A. 24 hours post-transfection, cells were also treated with either DMSO or 500 nM lapatinib for 24 hours.
Project description:This dataset consists of in situ HiC-seq data from a human oesophageal adenocarcinoma cell line (OE19). In total, the dataset includes 2 biological replicated samples. The Hi-C sample and library preparations were generated using Arima-HiC Kit (A510008, ARIMA Genomics) and Arima Library Prep module (A303011, ARIMA Genomics), respectively.