|
| Status |
Public on Aug 11, 2011 |
| Title |
VSMC AngII+CsA rep4 |
| Sample type |
RNA |
| |
|
| Source name |
Primary VSMC_AngII+CsA
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57BL/6
tissue origin: aorta
cell type: Primary vascular smooth muscle cells
|
| Treatment protocol |
Mouse VSMC were growth in DMEM medium supplemented with 20% heat-inactivated fetal bovine serum. The cultures were incubated with AngII with or without CsA at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was obtained with Trizol and Quiagen columns.
|
| Label |
Cy3
|
| Label protocol |
600 ng of total RNA, were reverse transcribed using T7 promoter primer and the Moloney murine leukemia virus (MMLV) reverse transcriptase (RT). cDNA was then converted to anti-sense RNA (aRNA) by using T7 RNA polymerase that amplifies target material and incorporates cyanine 3 (Cy3)-labeled CTP simultaneously.
|
| |
|
| Hybridization protocol |
1.65 micrograms of Cy3-labeled aRNA were hybridized for 17 hours at 65ºC in a Agilent hybridization oven (G2545A, Agilent Technologies) set to 10 rpm in a final concentration of 1x GEx Hybridization Buffer HI-RPM (Agilent Technologies). Arrays were washed and dried out using a centrifuge according to manufacturer's instructions (One-Color Microarray-Based Gene Expression Analysis, Agilent Technologies).
|
| Scan protocol |
Arrays were scanned at 5um resolution on an Agilent DNA Microarray Scanner (G2565BA, Agilent Technologies) using the default settings for 4x44k format one-color arrays.
|
| Data processing |
Raw data files from Feature Extraction were preprocessed with GeneSpring v10 (Agilent) : Raw signal values (in log2) were thresholded to 1 and arrays were normalized using quantiles (Bolstad et al. 2003).
|
| |
|
| Submission date |
Aug 10, 2011 |
| Last update date |
Aug 11, 2011 |
| Contact name |
Vanesa Esteban |
| E-mail(s) |
[email protected]
|
| Organization name |
Centro Nacional de Investigaciones Cardiovasculares (CNIC)
|
| Department |
Department of Vascular Biology and Inflammation
|
| Lab |
Regulation of Gene Expression in Vascular Endothelium
|
| Street address |
Melchor Fernández Almagro, 3.
|
| City |
Madrid |
| State/province |
Madrid |
| ZIP/Postal code |
28029 |
| Country |
Spain |
| |
|
| Platform ID |
GPL7202 |
| Series (1) |
| GSE31321 |
AngII-regulated genes sensitive to CsA in Vascular Smooth muscle cells. |
|
