title: Sample 18
organism: Homo sapiens
description: Protocols: ER-alpha positive BC cells: MCF-7 (ATCC HTB-22) were propagated in Dulbecco's modified Eagle medium (DMEM; Sigma-Aldrich) supplemented with 10% FBS (HyClone) and antibiotics: 100 U/ml penicillin, 100 mg/ml streptomycin, 250 ng/ml amphotericin-B. None ER-alpha positive BC cells: MCF-7 (ATCC HTB-22) were propagated in Dulbecco's modified Eagle medium (DMEM; Sigma-Aldrich) supplemented with 10% FBS (HyClone) and antibiotics: 100 U/ml penicillin, 100 mg/ml streptomycin, 250 ng/ml amphotericin-B. ChIP-Seq experiments were performed in three biological replicates in MCF-7 cells, in exponential growing condition. Cells were treated with ICI 182,780 10-7 M for 3 days and with EPZ004777 6.4 µM or DMSO for 6 days. ChIP-Seq experiments were performed in three biological replicates in MCF-7 cells, in exponential growing condition. Cells were treated with ICI 182,780 10-7 M for 3 days and with EPZ004777 6.4 µM or DMSO for 6 days. We used antibodies to ERα (HC-20) (Santa Cruz Biotechnology, SC-8002), Dot1l Antibody (Bethyl Laboratories, A300-954A), and rabbit IgG antiobody as negative control. Cells lysis, sonication and binding were performed as described by Serandour AA et al, 2013. After binding, beads washing, elution, reverse crosslinking and DNA extraction were then performed as described in Ambrosino et al, 2010. 1µl aliquot of DNA obtained was analysed to asses size distribution by Agilent Genomic DNA ScreenTape & Reagents using an Agilent 4200 TapeStation System (Agilent Technologies). The concentration of each DNA sample was determined by using Quant-IT DNA Assay Kit-High Sensitivity and a Qubit Fluorometer (Life Technologies). 10 ng of each purified ChIP DNA were used for sequencing libraries preparation with TruSeq ChIP Sample Prep Kit (Illumina Inc.).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-02-08T17:04:05Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-06-11T13:57:32Z
attributes.tag: External Id
attributes.value: SAMEA4722178
attributes.tag: INSDC center name
attributes.value: Laboratory of Molecular Medicine and Genomics
attributes.tag: INSDC first public
attributes.value: 2019-02-08T17:04:05Z
attributes.tag: INSDC last update
attributes.value: 2018-06-11T13:57:32Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6883:Sample 18
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: MCF-7
attributes.tag: cell type
attributes.value: mammary epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: invasive ductal carcinoma
attributes.tag: metastatic site
attributes.value: pleural effusion
attributes.tag: organism part
attributes.value: mammary gland
attributes.tag: sample name
attributes.value: E-MTAB-6883:Sample 18
attributes.tag: scientific_name
attributes.value: Homo sapiens