2024-12-01
ENA first public
ERC000011
ENA-CHECKLIST
Silesian University of Technology
INSDC center name
public
INSDC status
ERS18371144
SRA accession
E-MTAB-13865:MCF7_TGF_Ctr_3
Submitter Id
ArrayExpress
broker name
MCF7
cell_line
epithelial cell
cell_type
not collected
collection date
human
common name
Protocols: Non-tumorigenic human breast epithelial MCF10A cells (authenticated in 2016) were cultured in DMEM/F12 medium supplemented with 5% horse serum (BioWest, Nuaillé, France), 5 µg/mL insulin (Sigma-Aldrich, Saint Louis, MO, USA), 0.5 µg/mL hydrocortisone (Sigma-Aldrich), 20 ng/mL EGF (Sigma-Aldrich). Breast cancer MCF7 cells were cultured in DMEM/F12 medium supplemented with 10% Fetal Bovine Serum (EURx, Gdansk, Poland). Cells were seeded on culture plates the day before the start with TGFβ1 stimulation, at various densities (~80% at the time of cells harvesting and RNA isolation). Each day TGFβ1 was administered in fresh medium for 2 h and then the medium was changed to fresh one for another 22 h to obtain conditioned medium (CM); CM from each treatment day was used to treat other cells to induce potential bystander effect. Total RNA isolation was performed at appropriate time points. Total RNA was isolated using the Direct-ZolTM RNA MiniPrep Kit (Zymo Research) and digested with DNase I (Worthington Biochemical Corporation). For each experimental point, RNAs from three biological replicates were first tested by RT-qPCR for the efficiency of treatments. Libraries were generated with Illumina TruSeq mRNA stranded (poly-A selection) with a customized fragmentation protocol (custom fragmentation protocol: 90'C for 2 minutes).
description
adult
developmental stage
adenocarcinoma
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
breast
organism part
3
replicate
Homo sapiens
scientific_name
treated with TGFβ1 (10ng/ml)
stimulus
MCF7_TGF_Ctr_3
title