|
| Status |
Public on Aug 09, 2010 |
| Title |
Aorta_cMycON_2days_biological rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Whole aorta, Tamoxifen treated, 2 days
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Whole aorta
genotype: CBA-C57BL/6 pIns-c-MycERTAM
gender: female
age: 3-6 months
|
| Treatment protocol |
c-MycERTAM protein is activated in pancreatic beta cells of adult transgenic mice, by daily intraperitoneal (IP) administration of tamoxifen (TAM; Sigma-Aldrich, Dorset, UK) (1 mg/0.2 ml in peanut oil). Inactivation of c-MycERTAM protein was achieved following withdrawal of TAM. All protocols conformed with the Home Office (UK) laws and regulations, and transgenic mice were registered with the University of Warwick, Biological Sciences Committee for Genetically Modified. Mice were killed by cervical dislocation. The thoracic cavity was opened rapidly and saline was perfused through the mouse heart while the aorta was intact. All remaining blood was washed off the aorta before isolation.
|
| Growth protocol |
Pelengaris S, Khan M, Evan G (2002) Suppression of Myc-induced apoptosis in beta cells exposes multiple innate oncogenic properties of Myc sufficient to trigger immediate carcinogenic progression. Cell 109: 321-334.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The thoracic part of the aorta was snap-frozen in liquid nitrogen for RNA extraction. Frozen mouse aortas were homogenised and total RNA was extracted using RNEasy Plus Mini-Kit (Qiagen, Crawley, UK), according to the manufacturer’s protocol. RNA concentration was estimated using a spectrophotometer (ND-1000 Nanodrop, Labtech International Ltd, East Sussex, UK) prior to further quality control check by a bioanalyzer (2100 Bioanalyzer, Agilent Technologies, Cheshire, UK). For this purpose, n=3 mice were used per timepoint, each mouse was analysed individually without pooling of RNA .
|
| Label |
biotin
|
| Label protocol |
Biotin
|
| |
|
| Hybridization protocol |
A two-cycle protocol was performed at the beginning of our gene array analysis. 10µg double-amplified biotin-labelled cRNA were hybridised to Affymetrix MOE430 2.0 GeneChips (Affymetrix UK Ltd, High Wycombe, UK)) together with pre-labelled hybridisation controls as described in the Affymetrix GeneChip Expression Analysis Technical Manual.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneArray 2500 Scanner.
|
| Description |
Adult female mice, 3-6 months of age, inbred into CBA-C57Bl/6 background and maintained under barrier conditions, were treated with TAM for up to 7 days (Myc ON) and then monitored during recovery for more than 4 months (140 days) (Myc OFF).
|
| Data processing |
The data were intially analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 100. Further processing was performed by RMA normalization.
|
| |
|
| Submission date |
Mar 25, 2009 |
| Last update date |
Aug 09, 2010 |
| Contact name |
Yi-Fang Wang |
| Organization name |
Warwick University
|
| Department |
Biological Sciences
|
| Street address |
Gibbet Hill Road
|
| City |
Coventry |
| ZIP/Postal code |
CV4 7AL |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE15401 |
Expression data from mouse aorta during and following transient hyperglycaemia |
|
