|
| Status |
Public on Jul 05, 2013 |
| Title |
AdERbetasiERalphaE2 rep1 |
| Sample type |
RNA |
| |
|
| Source name |
MCF-7 cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF-7
cell type: breast adenocarcinoma
infected with: adenovirus carrying estrogen receptor beta (AdERb)
transfected with: siERalpha
treated with: 10 nM E2 (Sigma-Aldrich) for 24h
|
| Treatment protocol |
MCF-7 cells expressing endogenous ERalpha were infected with adenovirus carrying either estrogen receptor beta (AdERb) or no insert (Ad) at multiplicity of infection (moi) of 50. ERβ only cells were generated from these cells by knockdown of ERα in parental cells using the following siERα sequences from Dharmacon: forward, 5’-UCAUCGCAUUCCUUGCAAAdTdT-3’, and reverse, 5’-UUUGCAAGGAAUGCGAUGAdTdT-3’. siRNA experiments were performed as previously described, and resulted in knocknown of ERα mRNA and protein by greater than 95% (Chang et al, 2008). Briefly, cells were transfected with 20 nM siCtrl or siERα for 48 h after infection. Then cells were treated with 0.1% EtOH (Veh) or 10 nM E2 (Sigma-Aldrich) for 24h. All experiments were conducted with two or more replicates.
|
| Growth protocol |
MCF-7 cells were maintained in MEM media with 5% Calf serum
|
| 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 10 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 Affymetrix [HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner GCS 3000.
|
| Description |
Gene expression data from MCF-7 cells, AdenoERbeta infection 72h, siERalpha treatment 24h, 10 nM E2 for 24h
|
| Data processing |
The data were analyzed with GCOS2.1 using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 100.
|
| |
|
| Submission date |
Nov 16, 2012 |
| Last update date |
Aug 19, 2014 |
| Contact name |
Zeynep Madak Erdogan |
| Organization name |
UIUC
|
| Department |
FSHN
|
| Lab |
Zeynep Madak Erdogan
|
| Street address |
1201 S Goodwin Ave
|
| City |
Urbana |
| State/province |
IL |
| ZIP/Postal code |
61801 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (2) |
| GSE42347 |
Integrative genomics of gene and metabolic regulation by estrogen receptors α and β and coregulators [expression] |
| GSE42349 |
Integrative genomics of gene regulation by estrogen receptors and and coregulators |
|
