|
| Status |
Public on Dec 18, 2009 |
| Title |
LeftVentricleEndothelialCells_AntioxidantandExercised4 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Left ventricle endothelial cells, antioxidant supplemented and exercised
|
| Organism |
Rattus norvegicus |
| Characteristics |
tissue: heart
cell type: endothelial
strain: Wistar
Sex: male
age: 24wk old
|
| Biomaterial provider |
UQBR, University of Queensland
|
| Treatment protocol |
Antioxidant supplementation for 14 weeks:
1000IU vitamin E /kg diet
1.6g alpha lipoic acid /kg diet
Endurance training on treadmill for 14 wks
|
| Growth protocol |
Rats were housed 2-3 per cage,
maintained on a 12-12h light/dark cycle,
provided with rat chow and tap water ad libitum
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Endothelial cells were extracted from left ventricles using Dynabeads. Total RNA was then extracted using a Quigen Rneasy Micro Kit according to the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
cDNA was labeled using a SuperScript Plus Direct Labeling System according to the manufacturer's instructions.
|
| |
|
| Channel 2 |
| Source name |
Left ventricle endothelial cells, reference
|
| Organism |
Rattus norvegicus |
| Characteristics |
tissue: heart
cell type: endothelial
strain: Wistar
Sex: male
age: 24wk old
|
| Biomaterial provider |
UQBR, University of Queensland
|
| Treatment protocol |
N/A
|
| Growth protocol |
Rats were housed 2-3 per cage,
maintained on a 12-12h light/dark cycle,
provided with rat chow and tap water ad libitum
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Endothelial cells were extracted from left ventricle using Dynabeads. Total RNA was then extracted using a Quigen Rneasy Micro Kit according to the manufacturer's instructions.
|
| Label |
Cy5
|
| Label protocol |
cDNA was labeled using a SuperScript Plus Direct Labeling System according to the manufacturer's instructions.
|
| |
|
| |
| Hybridization protocol |
To 10µl sample, the following were added in order: 1µl of 10µg/µl poly dA, 8µl of 20x SSC, 20µl of deionised formamide and 1µl of 20% w/v SDS. After heat denaturing at 95oC for 5 min followed by equilibration at 45oC for 60 min 38µl of hybridization mixture hybridization mixture was added to prewarmed microarray slides. The hybridization was allowed to proceed for 22 hrs at 45oC in hybrization chambers saturated with 40µl of formamide/MilliQ.. After hybridsation slides were washed sequentially in the dark for in 1x SSC in 0.1% w/v SDS, 0.5x SSC and 0.2x SSC at 450C for 10 min each, dried and scanned immediately.
|
| Scan protocol |
Microarray slides were scanned using a high resolution Agilent scanner and scanned images were processed with ImaGene 6.0 software.
|
| Description |
N/A
|
| Data processing |
Signals of all arrays were normalized per spot and per chip using intensity-dependent Lowess fit normalization in GeneSpring.
|
| |
|
| Submission date |
Dec 17, 2008 |
| Last update date |
Apr 19, 2012 |
| Contact name |
Aya Matsumoto |
| E-mail(s) |
amatsumoto@hms.uq.edu.au
|
| Phone |
0733656983
|
| Organization name |
University of Queensland
|
| Department |
Hunan Movement Studies
|
| Street address |
Blair Drive
|
| City |
Brisbane |
| State/province |
Qld |
| ZIP/Postal code |
4072 |
| Country |
Australia |
| |
|
| Platform ID |
GPL1747 |
| Series (1) |
| GSE14044 |
Effects of exercise training and antioxidant supplementation on endothelial cell gene expression |
|
