|
| Status |
Public on Aug 09, 2012 |
| Title |
MEKi 4 days, biological rep5 |
| Sample type |
RNA |
| |
|
| Source name |
Melanoma allograft_AZD6244 100mg/kg 4 days
|
| Organism |
Mus musculus |
| Characteristics |
host strain: nude mice
injected with: iNRAS-475 mouse melanoma cells
tissue: Melanoma allograft
treated with: AZD6244 MEK inhibitor 100mg/kg for 4 days
|
| Treatment protocol |
Tumors were harvested from nude mice hosts. Skin was trimmed off of the tumors and a small piece (~50mg) of tumor parenchyma was placed into RNA Later preservation reagent, refrigerated overnight at 4C, then stored at -80C. Samples were later placed into Trizol after minimal thawing and homogenized using a hand-held electric tissue homogenizer.
|
| Growth protocol |
The iNRAS-475 mouse melanoma cell line was injected intradermally into nude mice which were fed 2mg/ml doxycycline water. Tumors were allowed to reach 200-500mm3 after 6 weeks. Mice were then treated with vehicle or 100mg/kg of the AZD6244 MEK inhibitor, or doxycyline was withdrawn from the diet.
|
| 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.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on a GeneChip Mouse Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
iNRAS-475 AZD d4 5
|
| Data processing |
Raw image files were converted to probe set data (.cel files) in Microarray Suite (MAS 5.0). The 18 probe set data files were normalized together, and expression values were determined, using the Robust Multi-chip Average (RMA) method as implemented in RMAExpress (Bolstad/Berkeley).
|
| |
|
| Submission date |
Aug 08, 2012 |
| Last update date |
Aug 09, 2012 |
| Contact name |
Lawrence N Kwong |
| E-mail(s) |
lkwong@mdanderson.org
|
| Organization name |
UT MD Anderson Cancer Center
|
| Street address |
1901 East Rd
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77054 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (2) |
| GSE39984 |
Comparison of the genetic extinction of NRAS to pharmacological MEK inhibition in an inducible mouse model of melanoma |
| GSE39987 |
Oncogenic NRAS Signaling Differentially Regulates Survival and Proliferation in Melanoma. |
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