|
| Status |
Public on Mar 29, 2008 |
| Title |
HSC3_3-siNTR1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Control siRNA in HSC3
|
| Organism |
Homo sapiens |
| Characteristics |
Control siRNA in HSC3
|
| Treatment protocol |
Transfection into HSC2 and HSC3 cells was performed using a final concentration of 20 nM siRNA and Lipofectamine RNAiMAX following the manufacturer's instructions.
|
| Growth protocol |
HSC2 and HSC3 were cultured in Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 Ham (DMEM/F-12) supplemented with 10% fetal bovine serum (FBS) in a humidified atmosphere containing 5% CO2 at 37°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Labeled cRNAs were generated from 500 ng of total RNA using Agilent's Low RNA Input Linear Amplification Kit (P/N 5188-5340).
|
| |
|
| Channel 2 |
| Source name |
NTSR1 siRNA in HSC3
|
| Organism |
Homo sapiens |
| Characteristics |
NTSR1 siRNA in HSC3
|
| Treatment protocol |
Transfection into HSC2 and HSC3 cells was performed using a final concentration of 20 nM siRNA and Lipofectamine RNAiMAX following the manufacturer's instructions.
|
| Growth protocol |
HSC2 and HSC3 were cultured in Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 Ham (DMEM/F-12) supplemented with 10% fetal bovine serum (FBS) in a humidified atmosphere containing 5% CO2 at 37°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
Labeled cRNAs were generated from 500 ng of total RNA using Agilent's Low RNA Input Linear Amplification Kit (P/N 5188-5340).
|
| |
|
| |
| Hybridization protocol |
Equal volumes of Cy5 sample and Cy3 sample were combined, mixed with 2x hybridisation buffer (Agilent Technoloiges) and hybridized to a Whole Rat Genome ologo array (G4131A, Agilent Technologies), for 17 hours at 60°C in an Agilent hybridization oven with rotation (Agilent Technologies).
|
| Scan protocol |
Scanned on an Agilent G2505B scanner and images were quantified using Agilent Feature Extraction Software (version 9.1).
|
| Description |
HSC3_3-siNTR1
|
| Data processing |
Data were extracted using Agilent's Feature Extraction software, version 9.1 (P/N G2565BA), Cy3 corresponding to the Processed Signal column of FE file, Cy5 corresponding to the rProcessedSignal column of FE file.
|
| |
|
| Submission date |
Mar 27, 2008 |
| Last update date |
Mar 28, 2008 |
| Contact name |
Satoya Shimizu |
| E-mail(s) |
smzsty@graduate.chiba-u.jp
|
| Organization name |
Chiba university
|
| Street address |
1-8-1 Inohana, Chuo-ku
|
| City |
Chiba |
| ZIP/Postal code |
260-8670 |
| Country |
Japan |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE10959 |
siNTSR1-treated HNSCC cell lines: Control vs. siNTSR1-treated HSC2 or HSC3 |
|
