|
| Status |
Public on Mar 03, 2017 |
| Title |
MCF-7_BPS_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Breast cancer cell line
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF-7
exposure time: 48h
experimental group: Bisphenol S
treatment: Bisphenol S
concentration (nm): 1500
|
| Treatment protocol |
MCF-7 cells were seeded into 96-well plates with DMEM medium containing 10% FBS at a density of 20,000 cells per well. After 24 h of steroid deprivation in hormone free medium (charcoal stripped FBS), the cells were stimulated with test substances for 48 h in triplicate in three independent experiments.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was performed using the Agencourt RNAdvance Cell V2 kit according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
Single Primer Isothermal Amplified (SPIA) cDNA was generated using the Ovation Pico WTA System V2 kit from Nugen, following the manufacturer's instructions. The SPIA cDNA was fragmented and Biotin-labelled using the Encore Biotin Module from Nugen according to the manufacturer instructions.
|
| |
|
| Hybridization protocol |
Hybridisation cocktails were prepared of the fragmented labelled-cDNA according to Nugen's recommendations and hybridised at 45°C overnight. The arrays were washed and stained using wash protocol FS450_0001 recommended for Affymetrix Human Transcriptome 2.0 Arrays on the GeneChip Fluidics station 450.
|
| Scan protocol |
Arrays were scanned using the GCS3000 7G scanner (Affymetrix) following manufacturer's protocols.
|
| Data processing |
CEL files were imported into Expression Console software (Affymetrix) and normalised using RMA-sketch (gene-level).
|
| |
|
| Submission date |
Aug 09, 2016 |
| Last update date |
Mar 06, 2017 |
| Contact name |
Robin Mesnage |
| E-mail(s) |
robinmes@gmail.com
|
| Phone |
+447482206767
|
| Organization name |
Kings College London
|
| Street address |
Guy's Hospital, 8th Floor, Tower Wing
|
| City |
London |
| ZIP/Postal code |
SE1 9RT |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL17586 |
| Series (1) |
| GSE85350 |
Bisphenol A alternatives can effectively substitute for estradiol in promoting cell growth through estrogen receptor alpha in human breast cancer cells |
|
