|
| Status |
Public on Nov 11, 2011 |
| Title |
hNHEPS_10 μM_6 h_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
hNHEPS human hepatocytes exposed to 10 μM VX 6 h after exposure
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Normal human primary hepatocytes isolated from non-transplantable donor tissue
time point: 6 h
dose: 10 μM
agent: VX (O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothiolate)
|
| Treatment protocol |
VX was added to cell culture at the specified concentrations and maintained for 1 h. Then VX was removed and the culture was washed 3X with fresh medium. The culture was then maintained at 37 °C with 5% CO2 until the indicated time points.
|
| Growth protocol |
Human hNHEPS hepatocytes were seeded in 6-well plates in media formulated by the manufacturer (Lonza, Walkersville, MD) and cultured at 37 °C under humidified 5% CO2 for 48 h before VX exposure.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the RNeasy Plus Mini Kit from Qiagen (Valencia, CA) following the manufacturer’s protocol, including an on-column DNase digestion.
|
| Label |
biotin
|
| Label protocol |
~0.1 μg of total RNA were labeled using the Affymetrix (Santa Clara, CA) GeneChip 3’ IVT Express Kit following the manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
Hybridization was carried out using the Affymetrix GeneChip Hybridization, Wash, and Stain Kit following the manufacturer’s protocols.
|
| Scan protocol |
The chips were scanned using an Affymetrix GeneChip Instrument System and the image (.DAT) files were processed using the Affymetrix GeneChip Operating Software (GCOS) to generate cell intensity (.CEL) files.
|
| Description |
gene expression data of hNHEPS human hepatocytes exposed to 10 μM VX 6 h after exposure
|
| Data processing |
The data were averaged and normalized in Partek Genomics Suite 6.5 (Partek, St. Louis, MO) using the robust multiarray analysis (RMA) algorithm.
|
| |
|
| Submission date |
Nov 10, 2011 |
| Last update date |
Nov 11, 2011 |
| Contact name |
Xiugong Gao |
| E-mail(s) |
xiugong.gao@amedd.army.mil
|
| Organization name |
Walter Reed Army Institute of Research
|
| Street address |
503 Robert Grant Ave
|
| City |
Silver Spring |
| State/province |
MD |
| ZIP/Postal code |
20910 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE33606 |
Gene expression changes in human hepatocytes exposed to VX (O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothiolate) |
|
