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Unknown,Transcriptomics,Genomics,Proteomics

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ChIP-Seq of human MCF-7 cells with ER-alpha and ER-beta anitbodies following estrogen treatment

ABSTRACT: We performed chromatin immunoprecipitation (ChIP) with overnight with antibodies against the C-terminus (HC-20, from Santa Cruz Biotechnology, Europe) or the N-terminus (anti-Estrogen Receptor 18-32, from SigmaAldrich, Italy) of human ER-alpha with or without estrogen treatment for 45 minutes on TAP-ER-beta cells, and the immunoprecipitated DNA was sequenced with the Illumina/Solexa Genome Analyzer. (Note: Raw data files not provided by submitter)

INSTRUMENT(S): Solexa 2G Genome Analyzer

ORGANISM(S): Homo sapiens

SUBMITTER: Alessandro Weisz

PROVIDER: E-MTAB-345 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Global analysis of estrogen receptor beta binding to breast cancer cell genome reveals an extensive interplay with estrogen receptor alpha for target gene regulation.

Grober Oli M V OM Mutarelli Margherita M Giurato Giorgio G Ravo Maria M Cicatiello Luigi L De Filippo Maria Rosaria MR Ferraro Lorenzo L Nassa Giovanni G Papa Maria Francesca MF Paris Ornella O Tarallo Roberta R Luo Shujun S Schroth Gary P GP Benes Vladimir V Weisz Alessandro A

BMC genomics 20110114

<h4>Background</h4>Estrogen receptors alpha (ERα) and beta (ERβ) are transcription factors (TFs) that mediate estrogen signaling and define the hormone-responsive phenotype of breast cancer (BC). The two receptors can be found co-expressed and play specific, often opposite, roles, with ERβ being able to modulate the effects of ERα on gene transcription and cell proliferation. ERβ is frequently lost in BC, where its presence generally correlates with a better prognosis of the disease. The identif ...[more]

PMID: 21235772

Similar Datasets

ChIP-Seq of human MCF-7 cells with ER-alpha anitbody following estrogen treatment

Project description:We performed chromatin immunoprecipitation (ChIP) with two polyclonal anti-ERalpha antibodies directed against the N- and C-terminus of the protein, with or without estrogen treatment for 45 minutes of MCF-7 cells, and the immunoprecipitated DNA was sequenced with the Illumina/Solexa Genome Analyzer.

2010-01-11 | E-MTAB-131 | biostudies-arrayexpress

MicroRNA profiling by array of human MCF-7 breast cancer cells with ER-beta tagged at the C-terminal or N-terminal and treated with 17-beta-estradiol

Project description:Human estrogen-responsive breast cancer cell line MCF-7 TET Off (MCF-7 wt) were used to produce stable clones expressing ER-beta tagged with TAP-tag respectively at the C-term and at the N-term (C-TAP-ER-beta and N-TAP-ER-beta) or expressing ER-alpha tagged (C-TAP-ER-alpha) as previously described.(C-TAP-ER-beta, N-TAP-ER-beta, C-TAP-ER-alpha) were treated with 10nM of17-beta-estradiol, or vehicle (ETOH). miRNA expression was analyzed on total RNA extracted before or after 6, 12, 24, and 72 hours hormonal stimulation. Total RNA was fluorescently labelled, amplified in triplicate, to be, than, pooled for the Hybridization.Each pool, were hybridized for 18 hours on Illumina v2 MicroRNA Expression BeadChips, and after scanning, analysis was performed with GenomeStudio v.2010.1 software, for quality control and miRNA expression analysis.

2011-07-29 | E-TABM-1055 | biostudies-arrayexpress

Formation of ER-lumenal intermediates during export of Plasmodium proteins containing transmembrane-like hydrophobic sequences

Project description:REX3RQLSE:Pf332:C-S11:DSLE is exported and is diffusely localised in the red blood cell cytoplasm (Fig 3A). Mutation of the PEXEL sequence (REX3AQLSE:Pf332:C-S11:DSLE), results in the protein being retained in the parasite (Fig 3C), and addition of a C-terminal SDEL sequence (REX3RQLSE:Pf332:C-S11:SDEL) also leads to retention of the protein within the parasite (Fig 3B). Given that REX3RQLSE:Pf332:C-S11:SDEL is retained within the parasite, this indicates that the C-terminus of the protein is in the ER lumen. To determine the location of the N-terminal end of this ER-retained protein, it was purified for tryptic digestion and analysis by mass spectrometry. The most N-terminal peptides retrieved corresponded to N-acetylated and non-acetylated SEPVVEEQDLK and SEPVVEEQDLKK. These peptides correspond to the expected N-terminal sequence following PEXEL cleavage by Plasmepsin V, indicating that the N-terminus of this protein is also in the ER-lumen. These data indicate that both N- and C-terminal ends of REX3RQLSE:Pf332:C-S11:SDEL are located within the ER lumen and consequently that in the context of this protein, the putative transmembrane segment of Pf332 has a propensity to translocate into the ER lumen rather than integrate into the ER membrane.

2023-03-28 | PXD036904 | Pride

Gene expression profile in breast cancer cells

Project description:Human estrogen-responsive breast cancer cell line MCF-7 wt were used to produce stable clones expressing ER-beta tagged with TAP-tag respectively at the C-term and at the N-term (Ct-ER-beta and Nt-ER-beta) as previously described. MCF7 wt and beta clone cells were cultured in steroid-free medium for 5 days and then were treated with 10nM of 17-beta-estradiol, or vehicle (ETOH). RNA was extracted after 2h, 4h and 8h of stimulation with 17-ß-estradiol 10 nM (+E2) or ethanol vehicle . Total RNA extracted by Ct-ER-beta and Nt-ER-beta cells were pooled (TAP-ER-beta). For mRNA expression profiling, 500 ng total RNA were reverse transcribed and used for synthesis of cDNA and biotinylated cRNA. Finally cRNA were hybridized for 18 hours on Illumina HumanHT-12 v3.0 BeadChips and after scanning, data analysis was performed.

2011-07-29 | E-TABM-1051 | biostudies-arrayexpress

ER, GR, and E2

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2013-07-01 | E-GEOD-46124 | biostudies-arrayexpress

Transcription profiling by array of human MCF-7 breast cell clones expressing ER-beta tagged with TAP-tag at the C-terminal or N-terminal

Project description:MCF-7 TET Off cells (MCF-7 wt) were used to produce stable clones expressing ER-beta tagged with TAP-tag respectively at the C-term and at the N-term (C-TAP-ER-beta and N-TAP-ER-beta) or expressing ER-alpha tagged (C-TAP-ER-alpha) as previously described.Cells were cultured in standard growth conditions, then were lysed and RNA was extracted using TRIzol method.Total RNA were fluorescently labelled, amplified and hybridized in triplicate (MCF7-TAP and C-TAP-ER-alpha) and in duplicate (C-TAP-ER-beta_A, C-TAP-ER-beta_B, N-TAP-ER-beta) for 18 hours on Illumina v2 MicroRNA Expression BeadChips and after scanning, analysis was performed with GenomeStudio v.2010.1 software, for quality control and miRNA expression analysis.

2011-10-11 | E-TABM-1053 | biostudies-arrayexpress

ER and GR crosstalk at the Genomic Level in mouse mammary cells

Project description:We report changes in ER and GR binding profiles genome-wide upon co-treatment with Dex and E2 when compared to Dex or E2 treatments alone. We examine ER and GR binding under four different treatments (unt, Dex, E2, and Dex + E2).

2013-07-01 | E-GEOD-46123 | biostudies-arrayexpress

Foxo3 Regulated Genes in G1E-ER-GATA-1 Cells

Project description:The ensemble of Foxo3-regulated genes in the erythroid G1E-ER-GATA-1 cell line was determined by knocking down Foxo3 using siRNA, and measuring genome wide transcription by microarray analysis G1E-ER-GATA-1 cells were treated with control or Foxo3-specific siRNA by nucleofection at t = 0 h and t = 24 h. At t = 24 h, cells were treated with M-CM-^_-estradiol to activate ER-GATA-1. RNA was harvested at t = 48 h and processed for microarray analysis.

2014-08-09 | E-GEOD-60137 | biostudies-arrayexpress

Transcription profiling by array of human MCF-7 cells with ER-beta tagged with TAP-tag at the C-term or N-term or expressing ER-alpha tagged

Project description:MCF-7 TET Off cells (MCF-7 wt) were used to produce stable clones expressing ER-beta tagged with TAP-tag respectively at the C-term and at the N-term (C-TAP-ER-beta and N-TAP-ER-beta) or expressing ER-alpha tagged (C-TAP-ER-alpha). All were grown in Dulbecco's modified Eagle's medium (DMEM). Then cells were lysed and RNA extracted were pooled. For mRNA expression profiling, 500 ng total RNA were reverse transcribed and used for synthesis of cDNA and biotinylated cRNA. Finally cRNA were hybridized for 18 hours on Illumina HumanHT-12 v3.0 BeadChips and after scanning, data analysis was performed.

2011-10-11 | E-TABM-1052 | biostudies-arrayexpress

GATA-1 and heme regulate the erythroid cell transcriptome.

Project description:Alas2 gene encodes the rate-limiting enzyme in heme biosynthesis. CRISPR/Cas9-mediated ablation of two Alas2 intronic cis-elements strongly reduced GATA-1-induced Alas2 transcription, heme biosynthesis, and GATA-1 regulation of other vital constituents of the erythroid cell transcriptome. Bypassing Alas2 function in Alas2 cis-element-mutant (double mutant) cells by providing its catalytic product 5-aminolevulinic acid (5-ALA) rescued heme biosynthesis and the GATA-1-dependent genetic network. We discovered a GATA factor- and heme-dependent circuit that establishes the erythroid cell transcriptome. G1E-ER-GATA-1 WT and double mutant cells were examined. Untreated WT, beta-estradiol-treated WT, beta-estradiol-treated double-mutant, and beta-estradiol/5-ALA-treated double-mutant cells were subjected to RNA-seq.

2016-01-07 | E-GEOD-74371 | biostudies-arrayexpress

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