|
| Status |
Public on Jun 01, 2007 |
| Title |
a4ERbeta1h-tet+E2(5)UHR(3)array99 T47D cells with inducible ERbeta |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
T-47D cells with inducible ERbeta construct
|
| Organism |
Homo sapiens |
| Characteristics |
T-47D breast cancer cell line
Tissue: breast cancer cell line
|
| Growth protocol |
Cells were added to 150 mm plates at a confluency of 40%; after one day, the normal medium was replaced by the medium described above, supplemented with 5% dextran coated charcoal treated FBS (DCCFBS). After 24h, 10 nM ICI 182,780 was added to the cultures and incubation proceeded for an additional 48h. For expression of ERb, tetracycline was removed 12 h before initiation of treatment with 17b-estradiol. At time 0 h the medium was changed to 0.5% DCCFBS and 10 nM of 17b-estradiol or equivalent volume of DMSO for mock-treated controls were added.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
25 ug each of sample total RNA and human universal reference RNA (Stratagene, La Jolla, CA, USA) were labeled with Cy5-conjugated dUTP and Cy3-conjugated dUTP (PerkinElmer, Boston, MA, USA), respectively, and hybridized to the arrays using protocols established by the Patrick O. Brown Laboratory (Stanford University, USA) accessible at http://cmgm.stanford.edu/pbrown/protocols/index.html.
|
| |
|
| Channel 2 |
| Source name |
Universal human reference RNA (Stratagene)
|
| Organism |
Homo sapiens |
| Characteristics |
Purchased from Stratagene
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
25 ug each of sample total RNA and human universal reference RNA (Stratagene, La Jolla, CA, USA) were labeled with Cy5-conjugated dUTP and Cy3-conjugated dUTP (PerkinElmer, Boston, MA, USA), respectively, and hybridized to the arrays using protocols established by the Patrick O. Brown Laboratory (Stanford University, USA) accessible at http://cmgm.stanford.edu/pbrown/protocols/index.html.
|
| |
|
| |
| Hybridization protocol |
Brown Lab protocol
|
| Scan protocol |
Scanned on an Axon 4000B scanner.
Images were quantified using GenePix Pro Software.
|
| Description |
induced and treated at 1 hour following E2 treatment
|
| Data processing |
Signal calculation: mean intensity minus median background and normalized by median value of all spot filtered genes.
|
| |
|
| Submission date |
Mar 06, 2007 |
| Last update date |
Mar 06, 2007 |
| Contact name |
Chin-Yo Lin |
| E-mail(s) |
chinyolin@byu.edu
|
| Phone |
801-422-6259
|
| Fax |
801-422-0519
|
| Organization name |
Brigham Young University
|
| Department |
Microbiology and Molecular Biology
|
| Street address |
753 WIDB
|
| City |
Provo |
| State/province |
UT |
| ZIP/Postal code |
84602 |
| Country |
USA |
| |
|
| Platform ID |
GPL4840 |
| Series (1) |
| GSE7206 |
Gene expression profiling in T-47D breast cancer cells following inducible expression of estrogen receptor beta |
|
