|
| Status |
Public on May 06, 2009 |
| Title |
cyto 4 |
| Sample type |
RNA |
| |
|
| Source name |
Cytokine treated HepG2 Cells
|
| Organism |
Homo sapiens |
| Characteristics |
hnf4 si rna treatment: untreated
cytokine treatment: treated
|
| Treatment protocol |
The inflammatory response in HepG2 cells was stimulated with a cytokine mixture containing 1 ng/ml of recombinant human IL-1β, 10 ng/ml of IL-6, and 10 ng/ml of TNF-α in serum-free medium for 18 h. Knock-down of HNF-4α in HepG2 cells was transfected with shRNA (HNF-4α shRNA and non-specific control shRNA) using the Nucleofector (Amaxa Biosystem) following the manufacturer's instructions. Twenty-four hours after transfection, the cells were placed in serum-free medium for 5 h and then either treated or untreated with cytokines for 18 h.
|
| Growth protocol |
HepG2 cells were grown in Dulbecco Modified Eagle Medium supplemented with penicillin (100 units/ml), streptomycin (100 ug/ml), and 10% heat-inactivated fetal bovine serum at 37° C in a humidified atmosphere with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNeasy Mini kit (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 ug total RNA
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA was hybridized for 16 hr at 45°C on the GeneChip Human Genome U133 A 2.0 (Affymetrix, Santa Clara, CA). GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using Affymetrix GeneArray Scanner 3000 7G Plus (Affymetrix, Santa Clara, CA).
|
| Description |
Gene Expression In Cytokine treated HepG2 Cells
|
| Data processing |
Microarray data were quantified and normalized using Affymetrix MicroArray Suite (MAS) 5.0
|
| |
|
| Submission date |
May 06, 2009 |
| Last update date |
May 08, 2009 |
| Contact name |
Peter A Burke |
| E-mail(s) |
peter.burke@bmc.org
|
| Phone |
617-414-8056
|
| Fax |
617-414-7398
|
| Organization name |
Boston University School of Medicine
|
| Department |
Surgery
|
| Lab |
Trauma
|
| Street address |
850 Harrison Avenue, Dowling 2 South
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02118 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE15991 |
Expression profile analysis of inflammatory response regulated by hepatocyte nuclear factor 4α |
|
