|
| Status |
Public on Feb 18, 2009 |
| Title |
MKO2_plusHGF |
| Sample type |
RNA |
| |
|
| Source name |
liver; c-Met knockout mouse; stimulated with HGF
|
| Organism |
Mus musculus |
| Characteristics |
Backgroud:C57/BL6, Gender:male, Age:8-10 weeks
|
| Treatment protocol |
For gene array analysis c-MetDhepa and c-MetloxP/loxP controls were stimulated for 2 hours with 2µg recombinant mouse HGF.Three animals per group were treated in parallel, before and after i.p. injection of recombinant HGF or NaCl.
|
| Growth protocol |
C-MetloxP/loxP animals were first crossed with albumin/AFP promoter/enhancer13 (Alfp-Cre) or albumin promoter-Cre transgenic animals (Alb-Cre). The Alfp-Cre animals express Cre-recombinase during embryonal development, while Alb-Cre mice start to express Cre 2 weeks after birth.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cryopreserved liver tissue by using peqGold RNAPure (PeqLab, Erlangen, Germany).
|
| Label |
biotin
|
| Label protocol |
The Affymetrix GeneChip RNA One cycle Amplification Kit was used to prepare labelled cRNA from 5 ug of total RNA. The protocol was conducted using the reagents provided by Affymetrix in the One-Cycle cDNA synthesis kit (P/N 900431), One-Cycle IVT labelling kit (P/N 90449) and GeneChip Sample Cleanup Module (P/N 900371). A detailed description can be found in the Genechip Expression Analysis Technical Manual, section 2 (Eukaryotic Sample and Array Processing), chapter 1 (Eukaryotic Target Preparation) (P/N 701025, revision 6).
|
| |
|
| Hybridization protocol |
Hybridisation of 10ug cRNA was done overnight for 16 hours at 45ºC in a Hybridisation Oven 640 (Affymetrix). The protocol is conducted as described in the Genechip Expression Analysis Technical Manual, section 2 (Eukaryotic Sample and Array Processing), chapter 2 (Eukaryotic Target Hybridization) (P/N 701027, revision 5).
|
| Scan protocol |
Arrays were scanned on an Affymetrix 3000 7G scanner, as described in the Genechip Expression Analysis Technical Manual, section 2 (Eukaryotic Sample and Array Processing), chapter 2 (Eukaryotic Arrays: Washing, Staining and Scanning (P/N 701028, revision 5).
|
| Description |
A85_10_plusHGF_MKO2
|
| Data processing |
Expression estimates were calculated using GCRMA, employing the empirical Bayes approach for background adjustment.
|
| |
|
| Submission date |
Dec 16, 2008 |
| Last update date |
Feb 18, 2009 |
| Contact name |
Guido Hooiveld |
| E-mail(s) |
guido.hooiveld@wur.nl
|
| Organization name |
Wageningen University
|
| Department |
Div. Human Nutrition & Health
|
| Lab |
Nutrition, Metabolism & Genomics Group
|
| Street address |
HELIX, Stippeneng 4
|
| City |
Wageningen |
| ZIP/Postal code |
NL-6708WE |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL7546 |
| Series (1) |
| GSE13992 |
c-Met confers protection against chronic liver tissue damage and fibrosis progression after bile-duct-ligation in mice |
|
