|
| Status |
Public on Jan 13, 2012 |
| Title |
hSFC E2 0.01nM 24h Cell 2 |
| Sample type |
RNA |
| |
|
| Source name |
human skin fibroblast cells, 24h, 0.01nM E2, Cell2
|
| Organism |
Homo sapiens |
| Characteristics |
congenital condition: hypospadias
treatment: 0.01 nM E2
cell type: Skin fibroblast
gender: Male
|
| Growth protocol |
hSFCs were maintained in DMEM/Ham's F-12 (048-29785, Wako, Osaka, Japan) containing 10% fetal bovine serum (FBS, Mediatech, Herndon, VA) and grown at 37°C in a 5% CO2 humidified incubator. For growth under steroid-free conditions, the cells were seeded in phenol red-free DMEM/Ham's F-12 (045-30665, Wako) containing 5% charcoal/dextran-treated FBS (Hyclone, Logan, UT). All the culture media contained 100 U/ml penicillin/streptomycin and 2 mmol/L L-glutamine (Mediatech, Herndon, VA).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from the culture cells after treated with chemicals for 24 h using an RNeasy Kit (Qiagen, Valencia, CA) in accordancewith the manufacturer’s instructions. Quantification and quality assessment of theisolated RNA samples were performed and verified using an Agilent Bioanalyzer2100 (Agilent Technologies, Palo Alto, CA)and aNanoDrop spectrophotometer (NanoDrop products, Wilmington, DE) in accordancewith the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
RNA was amplified into cRNA and labeled with Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technologies).
|
| |
|
| Hybridization protocol |
Sampleswere then hybridized to the G4851A SurePrint G3 Human GE 8x60Karray slides (60000 probes, Agilent Technologies).
|
| Scan protocol |
The arrays were scanned using Agilent Microarray Scanner (G2565BA, Agilent Technologies).
|
| Description |
Gene expression after 24h 0.01nM E2 treatment
|
| Data processing |
The scanned images were analyzed using the standard procedures described in Agilent Feature Extraction software 9.5.3.1 (Agilent Technologies). Data analysis was performed with GeneSpring GX12.0.2 (Agilent Technologies).
|
| |
|
| Submission date |
Jan 11, 2012 |
| Last update date |
Jan 20, 2012 |
| Contact name |
Sone Hideko |
| E-mail(s) |
hsone@nies.go.jp
|
| Phone |
81298502420
|
| Organization name |
National Institute for Environmental Studies
|
| Department |
Center for Environmental Risk Research
|
| Lab |
Environmental Exposure Research Section
|
| Street address |
onogawa16-2
|
| City |
tsukuba |
| State/province |
ibaraki |
| ZIP/Postal code |
305-8506 |
| Country |
Japan |
| |
|
| Platform ID |
GPL14550 |
| Series (1) |
| GSE35034 |
Gene expression of human skin fibroblast cells derived from hypospadias patients in response to low-dose bisphenol A exposure |
|
