Homo sapiens
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. MCF-10A (ATCC CRL-10317) was cultured in Mammary Epithelial Cell Growth Medium (MEGM BulletKit, Lonza, Catalog No. CC-3150) with added 100 ng/ml cholera toxin. Tamoxifen resistant MCF-7/LCC2 cells (referred to as LCC2) were kindly provided by Prof. Janne Lehtio, Department of Oncology-Pathology, Karolinska Institutet, Sweden. Tamoxifen resistant cells were maintained DMEM supplemented as described above with the addiction of 1 µM 4-hydroxytamoxifen (4-OHTAM) (Sigma-Aldrich) Total RNA extraction was performed by using TRIzol (Invitrogen) as previously described in Nassa et al, 2018. RNA concentration in each sample was assayed Quant-IT RNA Assay Kit-High Sensitivity and a Qubit Fluorometer (Life Technologies) and its quality and integrity assessed with Agilent RNA ScreenTape & Reagents using an Agilent 4200 TapeStation System (Agilent Technologies). mRNA sequencing experiments, indexed libraries were prepared using 1 µg of total RNA from MCF-7 treated with 4-OHT 10-7 M or vehicle for 3 days, with TruSeq Stranded Total mRNA Sample Prep Kit (Illumina Inc.)
ENA-FIRST-PUBLIC
2019-05-17T04:02:21Z
ENA-LAST-UPDATE
2018-06-11T12:01:54Z
External Id
SAMEA4721925
INSDC center name
Laboratory of Molecular Medicine and Genomics
INSDC first public
2019-05-17T04:02:21Z
INSDC last update
2018-06-11T12:01:54Z
INSDC status
public
Submitter Id
E-MTAB-6892:Sample 2
broker name
ArrayExpress
cell line
MCF-7/LCC2
cell type
mammary epithelial cell
common name
human
disease
invasive ductal carcinoma
metastatic site
pleural effusion
organism part
mammary gland
phenotype
tamoxifen resistant
sample name
E-MTAB-6892:Sample 2
scientific_name
Homo sapiens