|
| Status |
Public on Sep 19, 2013 |
| Title |
RJ17-3 |
| Sample type |
RNA |
| |
|
| Source name |
endothelial cells
|
| Organism |
Mus musculus |
| Characteristics |
tissue: pulmonary microvascular
genotype: wild type
age: 4–6-weeks-old
gender: female
background strain: C57BL/6
treatment: vehicle
|
| Treatment protocol |
For gene expression study, PMVEC cells were treated with recombinant IL-4 (rIL-4, 50 ng/mL, Sigma-Aldrich), or vehicle control for 24h (n=3).
|
| Growth protocol |
WT and IL-4 KO (C57BL/6 background; male, 8–12 weeks old) were were placed in a Plexiglas chamber maintained at 10% O2 (hypoxic group) or in a chamber open to room air (normoxic group) for 4 days (d) with a 12:12-hours (h) light-dark cycle. After exposure to hypoxia or normoxia, mice were sacrificed and the lungs were harvested for RNA extraction. Mouse PMVEC were isolated from 4–6-week-old female C57BL/6 mice. Cells were incubated at 37°C in a humidified atmosphere of 5% CO2 and were used at passages 3–5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total lung RNA was isolated from each animal or from PMVEC and extracted using the Trizol Reagent method (Invitrogen). Additional purification was performed on RNAeasy columns (Qiagen, Valencia, CA). The quality of total RNA samples was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA).
|
| Label |
biotin
|
| Label protocol |
Total RNA was labled with the Ambion TotalPrep RNA Amplification Kit according to manufacturers suggested protocols.
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
vehicle
|
| Data processing |
A single intensity (expression) value for each Illumina probe on the array was obtained using Illumina BeadStudio software with standard settings and no background correction. The expression values for all the probes for each sample were scaled to have median 256 (2^8) and then log (base 2) transformed before performing statistical analysis.
|
| |
|
| Submission date |
Sep 19, 2012 |
| Last update date |
Sep 19, 2013 |
| Contact name |
chris cheadle |
| E-mail(s) |
[email protected]
|
| Phone |
4105505984
|
| Organization name |
JHU
|
| Department |
SOM
|
| Lab |
JHBMC Genomics Core
|
| Street address |
5200 Eastern Ave
|
| City |
baltimore |
| State/province |
MD |
| ZIP/Postal code |
21224 |
| Country |
USA |
| |
|
| Platform ID |
GPL6885 |
| Series (1) |
| GSE40999 |
Th2 cytokine IL-4 is critical in initiating pulmonary vascular inflammation under hypoxic conditions and induces pulmonary vascular endothelial cell activation. |
|
