|
| Status |
Public on Oct 07, 2011 |
| Title |
siRNA treated, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Insulinoma 832/13 INS-1 cell line
|
| Organism |
Rattus norvegicus |
| Characteristics |
passage: between passages 7 and 25
cell line: Insulinoma 832/13 INS-1 cell line
treatment: COUP-TFII knockdown
|
| Treatment protocol |
COUP-TFII depletion experiments in the 832/13 INS-1 cell line were performed as described previously (Perilhou A, Tourrel-Cuzin C, Kharroubi I, Henique C, Fauveau V, Kitamura T, Magnan C, Postic C, Prip-Buus C, Vasseur-Cognet M (2008). The transcription factor COUP-TFII is negatively regulated by insulin and glucose via FoxO1 and ChREBP controlled pathways. . Mol Cell Biol 28: 6568-6579). Briefly, cells were transfected with COUP-TFII siRNA or a scrambled siRNA using a Amaxa device following manufacturer instructions.
|
| Growth protocol |
The rat insulinoma 832/13 INS-1 cell line was used between passages 7 and 25. Cells were cultured at 5% CO2–95% air at 37°C in RPMI 640 medium containing 11 mM D-glucose supplemented with 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin-streptomycin, 10 mM HEPES, 1 mM sodium pyruvate (Invitrogen), and 50 μM β-mercaptoethanol (Invitrogen) (INS-1 medium).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted and purified from cultured cells using the RNA-Plus reagent according to the manufacturer’s instructions (Q-BIOgene).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol starting from 1 ug total RNA (Expression Analysis Technical Manual, 701025 Rev5, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA was hybridized for 16 hr at 45C on 230 2.0 rat Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the affymetrix 3000 genescanner
|
| Data processing |
The data were 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 set to 150
|
| |
|
| Submission date |
Jul 08, 2011 |
| Last update date |
Oct 07, 2011 |
| Contact name |
Leentje Van Lommel |
| Organization name |
KULeuven
|
| Department |
Molecular cell biology
|
| Lab |
gene expression unit
|
| Street address |
Herestraat 49 bus 901
|
| City |
Leuven |
| ZIP/Postal code |
3000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL1355 |
| Series (1) |
| GSE30526 |
COUP-TFII Controls Mouse Postnatal Pancreatic β-Cell Mass through GLP-1 β-Catenin |
|
