|
| Status |
Public on Dec 09, 2006 |
| Title |
MCF7_dose_EGF_0.5nM_10min |
| Sample type |
RNA |
| |
|
| Source name |
breast epithelial cell, adherent, EGF, 0.5nM, 10min
|
| Organism |
Homo sapiens |
| Characteristics |
Cell line: MCF-7 (Obtained from ATCC (American Type Culture Collection)), Organ: mammary gland; breast, Cell type: epithelial, Disease: adenocarcinoma, Derived from metastatic site: pleural effusion, Age: 69 years adult, Gender: female, Ethnicity: Caucasian
|
| Treatment protocol |
Before treatments, medium were changed with serum-free medium again and incubated at 37C for 2 hours. Followed by incubation, added each ligand and incubated for each time.
|
| Growth protocol |
Prior to the growth factor (GF) treatment, cells were serum starved for 16-18 hours with serum-free DMEM medium and incubated at 37C/5% CO2. The medium was again changed 2 hr prior to the GF treatment.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol reagent (Invitrogen) and then purified using QIAGEN Rneasy kit.
|
| Label |
biotin
|
| Label protocol |
Standard Affymetrix Protocols (Affymetrix GeneChip Expression Analysis Technical Manual) was used. Complete protocal can be found manufacturer's web site (http://www.affymetrix.com/support/technical/manual/expression_manual.affx) (section2, chapter1-p13-18, & 2.1.p32-33,p36-40)
|
| |
|
| Hybridization protocol |
Standard Affymetrix Protocols (Affymetrix GeneChip Expression Analysis Technical Manual) was used. Complete protocal can be found manufacturer's web site (http://www.affymetrix.com/support/technical/manual/expression_manual.affx) (section2, 2.2.p3-8 & 2.3.p3-14)
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 with default parameter.
|
| Description |
Early gene expression (up to 90min) induced by growth hormone stimulation.
|
| Data processing |
The data were analyzed with GeneChip Operating System (GCOS) (ver.1.2) using Affymetrix default analysis settings and global scaling as normalization method. The Single-Array Expression Analysis function was used and the trimmed mean target intensity of each array was arbitrarily set to 500.
|
| |
|
| Submission date |
Dec 06, 2006 |
| Last update date |
Dec 08, 2006 |
| Contact name |
Mariko Okada |
| E-mail(s) |
marikoh@rcai.riken.jp
|
| Organization name |
RIKEN RCAI
|
| Lab |
Laboratory for Cellular Systems Modeling
|
| Street address |
W518, 1-7-22, Suehiro-cho, Tsurumi-ku
|
| City |
Yokohama |
| ZIP/Postal code |
230-0045 |
| Country |
Japan |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
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