|
| Status |
Public on Aug 22, 2013 |
| Title |
Lam3_1.0_a |
| Sample type |
RNA |
| |
|
| Source name |
Huh7 cell, IFNL3 treated,1h
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Huh7
treatment: Lambda3
time: 1
|
| Treatment protocol |
Huh7 cells were seeded into 6-well plates at the density of 3✕105 cells/well and cultured overnight before stimulation with either 500 U/ml of IFN-α or IFN-β, or 10 ng/ml of IFN-λ1, IFN-λ2, or IFN-λ3.
|
| Growth protocol |
Human Huh7 hepatoma cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with the addition of 10% heat-inactivated fetal bovine serum, 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 1 mM sodium pyruvate, 1X MEM non-essential amino acids and 10 mM HEPES buffer (Invitrogen)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was harvested and isolated at 0.5, 1, 2, 4, 6, 12 and 24 hours post-incubation using RNeasy Mini Kit (Qiagen)
|
| Label |
biotin
|
| Label protocol |
Preparation of cDNA, cRNA, and labeling were carried out by the Yale Center for Genomic Analysis using the standard Illumina protocols.
|
| |
|
| Hybridization protocol |
1.5 µg of cRNA was combined with hybridization controls and hybridized to the array for 16 hours in a hybridization oven at 58°C. The chip was then washed and stained with streptavidin-Cy3.
|
| Scan protocol |
The array was scanned using the Illumina BeadArray reader.
|
| Description |
Human Huh7 cells treated with 10ng/ml of IFN Lambda3 for 1 hours
|
| Data processing |
Microarray data from the validation cohort underwent quantile normalization using the Beadarray package in bioconductor.
|
| |
|
| Submission date |
Jun 28, 2013 |
| Last update date |
Aug 22, 2013 |
| Contact name |
Christopher Bolen |
| E-mail(s) |
cbolen1@gmail.com
|
| Organization name |
Yale University
|
| Department |
Pathology
|
| Lab |
Steven Kleinstein
|
| Street address |
300 George Street, Suite 505
|
| City |
New Haven |
| State/province |
CT |
| ZIP/Postal code |
06511 |
| Country |
USA |
| |
|
| Platform ID |
GPL10558 |
| Series (1) |
| GSE48400 |
Dynamic expression profiling of type I and type III interferon-stimulated hepatocytes. |
|
