|
| Status |
Public on Nov 16, 2009 |
| Title |
Endothelial Cells, High Shear Stress, 15 dyne/cm2, Replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Endothelial Cells, High Shear Stress, 15 dyne/cm2, Replicate 2
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Human Umbilical Vein Endothelial Cells
|
| Treatment protocol |
HUVEC were mounted in a parallel plate flow chamber and exposed to one of four conditions for 24 hours.
|
| Growth protocol |
HUVEC (passage 3-5) were grown to confluence on glass slides.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Immediately after exposure to shear stress total RNA was extracted using TRI-zol (Invitrogen); RNA was further purified with DNase (Qiagen) and RNeasy MinElute Cleanup Kit (Qiagen), according to manufacturers’ instructions. RNA quantity and quality were assessed by loading 1 ml of sample RNA onto a RNA 6000 Nano chip (Agilent) and analyzing with an Agilent 2100 Bioanalyzer to determine the amount and integrity of RNA recovered. Samples having an RNA Integrity Number of 9.5 or greater were used for microarray experiments.
|
| Label |
Cy3
|
| Label protocol |
RNA was linearly amplified to cRNA using Cy3 labeled CTP and a Low RNA Input Fluorescent Linear Amplification Kit (Agilent) according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Cy3-labeled cRNA for each sample was hybridized to a 44k Whole Human Genome Microarray (Agilent) according to the manufacturer’s instructions.
|
| Scan protocol |
Slides were dried under nitrogen and scanned on an Agilent DNA Microarray Scanner.
|
| Description |
Gene expression after exposure to high shear stress for 24 hours.
|
| Data processing |
The scanned images were amalyzed with Feature Extraction Software from Agilent and data was further analyzed using Genespring GX.
|
| |
|
| Submission date |
Jun 18, 2009 |
| Last update date |
Nov 16, 2009 |
| Contact name |
Marcie R Williams |
| E-mail(s) |
marcie@gatech.edu
|
| Organization name |
Georgia Institute of Technology
|
| Department |
Biomedical Engineering
|
| Lab |
Dr. Larry McIntire
|
| Street address |
2300 Country Walk 3427
|
| City |
Snellville |
| State/province |
GA |
| ZIP/Postal code |
30039 |
| Country |
USA |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE16706 |
Endothelial Responses to Atheroprone Flow |
|
