|
| Status |
Public on May 01, 2011 |
| Title |
MCF7 silenced Estrogen receptor, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
MCF7 silenced Estrogen receptor alpha (PII)
|
| Organism |
Homo sapiens |
| Characteristics |
knockdown: estrogen receptor alpha KD
cell line: MCF7
|
| Treatment protocol |
Endocrine independent cell line pII was established by transfection of MCF7 with ER directed shRNA plasmid. Another transfected cell line designated E2 was established that had retained sensitivity to estrogen and tamoxifen (presumably as a result of failure to produce siRNA) was used as an additional control.
|
| Growth protocol |
MCF7 derived from the ATCC (American Type Culture Collection, (VA, USA) were maintained at 37˚C in a humidified atmosphere of 5% CO2 in DMEM supplemented with 10% foetal bovine serum (FBS), 600 μg/ml L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin and 6 ml/500 ml 100 x non-essential amino acids (Invitrogen, CA, USA).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on Human U133 Plus 2 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix Scanner 3000 7G
|
| Description |
Gene expression data from MCF7 breast cancer cell line with silenced Estrogen receptor
|
| Data processing |
The data were analyzed with Partek Genomics Suite V 6.5 for RMA background correction, quantile normalisation with GC content adjustment
|
| |
|
| Submission date |
Feb 23, 2011 |
| Last update date |
May 01, 2011 |
| Contact name |
fahd Al-Mulla |
| E-mail(s) |
[email protected]
|
| Phone |
+96524986233
|
| Organization name |
Kuwait University
|
| Department |
Pathology
|
| Lab |
Molecular pathology
|
| Street address |
Faculty of medicine
|
| City |
Kuwait |
| ZIP/Postal code |
13110 |
| Country |
Kuwait |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE27473 |
Expression data from MCF7 cell line after silencing of Estrogen receptor |
|
