|
| Status |
Public on Apr 02, 2012 |
| Title |
MEFS p66KO H2O2 induced,biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
MEFS p66KO H2O2 induced
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Primary murine embryonic fibroblasts (MEFs)
strain: C57Bl/6
genotype: p66 knock-out (p66-/-)
age: 13.5-day-old embryos
|
| Treatment protocol |
Early passage (p.2) MEFs were washed twice with PBS1X solution, and then incubated in DMEM media containing 400 μM of H2O2 or 0.25μg/ml of Doxorubicin (SIGMA) at 37°C for four hours.
|
| Growth protocol |
Primary murine embryonic fibroblasts (MEFs) were isolated from 13.5-day-old embryos according to standard procedures from WT, p53KO and p66ShcKO mice. MEFs were cultured in DMEM supplemented with 10% fetal bovine calf serum, 100µg/ml streptomycin, 100 µg/ml penicillin FBS, L-glutamine (2 mM) and incubated at 37°C in a humidified atmosphere containing 5% CO2 and normal oxygen tension (20%). For all the experiments MEFs at early passages (p.2) were used. Treatment: Early passage (p.2-5) MEFs were washed twice with PBS1X solution, and then incubated in DMEM media containing 400 μM of H2O2 or 0.25μg/ml of Doxorubicin (SIGMA) at 37°C for four hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard Affymetrix 3' IVTprotocol from 100 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation,6 ug of cDNA were hybridized for 16 hr at 45C on GeneChip MOE430PM strip array. GeneChip strip was washed and stained in the Affymetrix GeneAtlas instrument .
|
| Scan protocol |
GeneChip strip images were acquired using the GeneAtlas Imaging Station.
|
| Description |
Gene expression data from MEFs passage 2
GeneChip MOE430PM strip arrays on Affymetrix GeneAtlas instrument
|
| Data processing |
The data were analyzed with Partek Genomics Suite v 6.5 using RMA algorithm.
|
| |
|
| Submission date |
Apr 06, 2011 |
| Last update date |
Apr 02, 2012 |
| Contact name |
Simone Paolo Minardi |
| E-mail(s) |
simone.minardi@ifom-ieo-campus.it
|
| Phone |
+39 02 574303703
|
| Organization name |
Cogentech
|
| Department |
Microarray and NGS Unit
|
| Street address |
Via Adamello
|
| City |
Milano |
| State/province |
Italy |
| ZIP/Postal code |
20139 |
| Country |
Italy |
| |
|
| Platform ID |
GPL11180 |
| Series (2) |
| GSE28416 |
Expression data from mouse embryo fibroblast (MEFS) |
| GSE28418 |
Expression data from mouse tissues and MEFs: insights into the physiological activation of p53-p66Shc pathway |
|
