|
| Status |
Public on Dec 31, 2016 |
| Title |
Human Mammary Epithelial Cells_Rheb_transfected_2 |
| Sample type |
RNA |
| |
|
| Source name |
HMECs_Rheb_transfected
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Human Mammary Epithelial cells (HMECs)
genotype/variation: over-expressing Rheb oncogene
|
| Treatment protocol |
Cells were brought to quisecence by serum starvation and treated with adenovirus to over-express the Rheb oncogene (10 replicates) or GFP (10 replicates). Eighteen hours after infection, activation of oncogenes and secondary targets were confirmed by western blotting.
|
| Growth protocol |
Mammary epithelial cells were isolated from a breast reductive surgery at Duke University and grown at 37 degrees C in mammary epithelial cell growth medium (MEGM) containing a MEGM Bullet Kit, human transferrin and isopenterol.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using a Qiagen Rneasy kit.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 3-4 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
The resulting biotin-labeled RNA samples were then hybridized with Affymetrix U133A 2.0 GeneChip arrays according to the manufacturer's instructions (Affymetrix, Santa Clara, CA).
|
| Scan protocol |
GeneChips were scanned using the Gene Array Scanner (Affymetrix) following the manufacturer’s instructions (Affymetrix, Santa Clara, CA).
|
| Description |
35203_Rheb2
|
| Data processing |
The data were analyzed using the R statistical platform with the Bioconductor affymetrix package [R version 2.13.1 (2011-07-08) and Bioconductor version 2.9].
|
| |
|
| Submission date |
Sep 27, 2013 |
| Last update date |
Dec 31, 2016 |
| Contact name |
David Hsu |
| Organization name |
Duke University
|
| Department |
Medical Oncology
|
| Street address |
905 S. LaSalle St. Rm 3008
|
| City |
Durham |
| State/province |
NC |
| ZIP/Postal code |
27710 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (2) |
| GSE51242 |
A Molecular Profile of Colorectal Cancer to Guide Therapy [HMEC] |
| GSE51244 |
A Molecular Profile of Colorectal Cancer to Guide Therapy |
|
