|
| Status |
Public on Nov 30, 2011 |
| Title |
Liver Nrf2 knock-out High Fat Diet, replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Liver Nrf2 knock-out 6 months High Fat Diet, replicate 3
|
| Organism |
Mus musculus |
| Characteristics |
treatment: high fat diet
tissue: liver
genotype: Nrf2 KO
gender: male
genetic background: C57BL/6J
|
| Treatment protocol |
Liver from wild type and Nrf2-KO mice was excised after perfusion with ice-cold saline and then it was immersed in RNA later solution (Ambion,Foster City, CA).
|
| Growth protocol |
Mice were housed in the animal facility in the University of Patras Medical School in temperature-, light- and humidity-controlled rooms with a 12-h light/dark cycle.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using a guanidinium thiocyanate based method using the Trizol reagent (Invitrogen, Carlsbad,CA) and was further purified using the Rneasy minikit (Qiagen,Hilden,Germany) following the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
Labelling of total RNA was performed in Atlas Biolabs (Berlin, Germany) using the following kit: Low Input Quick Amp Labeling Kit, One Color (#5190-2305), Agilent Technologies, using 100 ng of total RNA as starting material, according to the manufacturer´s instructions (Manual One-Color Microarray-Based Gene Expression Analysis, Low Input Quick Amp Labeling, Version 6.5, Mai 2010, Agilent Technologies). cRNA synthesis was regarded successful provided that =1.65 μg of cRNA with a Cy3-incorporation rate =8.0 pmol/μg cRNA were synthesized.
|
| |
|
| Hybridization protocol |
Hybridization was performed in Atlas Biolabs (Berlin,Germany) as follows;1.65 μg of Cy3-labelled cRNA were fragmented according to the manufacturer´s instructions, and 1.425 μg of fragmented cRNA were hybridized (conc: 15 ng/μl). Hybridization conditions: 65°C, 10 rpm, 17 h; in an Agilent hybridization oven
|
| Scan protocol |
Scanning was performed using an Agilent DNA Microarray Scanner (Model G2505C) and Agilent´s Scan Control Software (version A.8.3.1) in Atlas Biolabs (Berlin,Germany).
|
| Description |
Gene expression after 6 months on high-fat diet
|
| Data processing |
Primary data analysis was performed using Agilent´s Feature Extraction Software (version 10.7.3.1) in Atlas Biolabs (Berlin,Germany).
|
| |
|
| Submission date |
Nov 09, 2011 |
| Last update date |
Nov 30, 2011 |
| Contact name |
Dionysios Chartoumpekis |
| E-mail(s) |
dchart@upatras.gr
|
| Organization name |
Lausanne University Hospital CHUV
|
| Department |
Service of Endocrinology Diabetes and Metabolism
|
| Street address |
Ave de la Sallaz 8
|
| City |
Lausanne |
| ZIP/Postal code |
1011 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE33575 |
Hepatic gene expression profiles after long-term high-fat diet feeding in wild type and Nrf2 knock-out mice. |
|
