|
| Status |
Public on Mar 01, 2007 |
| Title |
MASMC_Hey2_KO_PDGF_T60 |
| Sample type |
RNA |
| |
|
| Source name |
MASMC Hey2 KO cells treated with PDGF
|
| Organism |
Mus musculus |
| Characteristics |
Genetic background: Mixed C57BL/6 and 129SvJ
Passage: 10-20
|
| Treatment protocol |
Mouse aortic smooth muscle cells were isolated from adventitia stripped, explanted aortas by mincing followed by enzymatic digestion. Cells were plated onto fibronectin coated dishes and cultured in DMEM with serum. Purity was checked by immunostaining for sm alpha actin and calponin.
|
| Growth protocol |
For PDGF induction, cells were grown to 95% confluency on 150 mm plates, washed 4 times with phosphate-buffered saline, and serum starved with DMEM supplemented with 0.4% FCS. After 48 hours in a 37¡/5% CO2 humidified incubator, the cells were washed four times with phosphate buffered saline and then either treated with DMEM or DMEM supplemented with 5 ng/ml human PDGF-BB. Cells were then incubated in the humidified incubator for 60 minutes.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After the appropriate length of treatment, the cells were washed one time with phosphate buffered saline and RNA was harvested with the Qiagen RNeasy protocol according to the manufacturerÕs instructions (Qiagen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 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 GeneChip Drosophila Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix Model 3000 scanner
|
| Description |
Gene expression data from Hey2 KO and WT MASMCs treated with PDGF for varying amounts of time.
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) 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 |
Dec 14, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Michael T Chin |
| E-mail(s) |
[email protected]
|
| Organization name |
University of Washington
|
| Department |
Medicine
|
| Street address |
815 Mercer Street
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98109 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE6526 |
Expression time course data HEY2 KO and WT MASMC treated with PDGF |
|
| Relations |
| Reanalyzed by |
GSE119085 |
