|
| Status |
Public on Aug 23, 2007 |
| Title |
HMVEC_SLx2119_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Human Microvascular Endothelial Cells, SLx-2119 treated 24 h
|
| Organism |
Homo sapiens |
| Characteristics |
Primary culture of human lung microvascular endothelial cells (CC-2527)
|
| Biomaterial provider |
Cambrex
|
| Treatment protocol |
HMVEC at passage 7 were plated in 6 cm culture dishes with 3 ml culture medium, at a density of 1,000,000 cells/dish. After 2 days (confluence 90%) the cells were incubated for 24 hours in 3 ml culture medium containing 40 µM SLx-2119 (small molecule ROCK-2 inhibitor).
|
| Growth protocol |
Human microvascular endothelial cells (HMVEC;CC-2527, Cambrex) were cultured in EGM-2 MV BulletKit medium, containing endothelial cell basal medium-2, human epidermal growth factor (hEGF), hydrocortisone, human basal fibroblast growth factor (hFGF-B), vascular endothelial growth factor, recombinant insulin-like growth factor 1, ascorbic acid, gentamicin, amphotericin-B, and 5% fetal bovine serum (FBS) and maintained at 37 ºC in a humidified atmosphere of 5% CO2 and 95% air.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
24 hours after treatment with SLx-2119, cells were harvested and total RNA was isolated using Ultraspec RNA isolation reagent (Biotecx Laboratories), according to the manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
200 ng of total RNA was reverse transcribed and amplified overnight with T7 RNA polymerase and labeled with biotin following the manufacturer’s protocol
|
| |
|
| Hybridization protocol |
1.5 μg of biotin-labeled aRNA was hybridized to Illumina Human Ref-6 v1 BeadChips at 58 °C overnight. BeadChips were incubated with Cy3 streptavidin and washed according to the manufacturer’s instructions.
|
| Scan protocol |
The hybridized BeadChips were scanned by Illumina BeadScan confocal scanner and analyzed by Illumina’s BeadStudio v1.5.1.3. aRNA synthesis, hybridization and scanning were performed by the Microarray Core Laboratory of the University of Texas Health Science Center.
|
| Description |
This sample is number 1 of 3 repeats.
Please note: The background of this array was 5 times higher than the background of the other arrays in the series. After background subtraction, values are still more than twice as high as values of the other arrays. We therefore excluded this sample from our subsequent analyses.
|
| Data processing |
Initial analysis was done using Illumina’s Beadstudio v1.5.1.3. The array was normalized to sample GSM214810 by rank-invariant method.
|
| |
|
| Submission date |
Aug 03, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Marjan Boerma |
| E-mail(s) |
mboerma@uams.edu
|
| Phone |
501-686-6882
|
| Organization name |
University of Arkansas for Medical Sciences
|
| Street address |
4301 West Markham, Slot 725
|
| City |
Little Rock |
| State/province |
AR |
| ZIP/Postal code |
72205 |
| Country |
USA |
| |
|
| Platform ID |
GPL2507 |
| Series (1) |
| GSE8686 |
Gene expression profiling in three primary human cell lines treated with atorvastatin or an inhibitor of Rho kinase |
|
