|
| Status |
Public on Dec 07, 2007 |
| Title |
Untreated Control at 3 hr (C_1.1 3) |
| Sample type |
RNA |
| |
|
| Source name |
Human Dermal Fibroblast
|
| Organism |
Homo sapiens |
| Characteristics |
Cell line
|
| Treatment protocol |
Cell were treated with 10 nM of marinobufagenin (MBG) for the indicated times follows by RNA isolation.
|
| Growth protocol |
Human skin fibroblasts, obtained from Cambrex, were cultured at 37oC in 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Qiagen's Rneasy column according to manufacturer's specification.
|
| Label |
Cy5
|
| Label protocol |
cDNA were synthesized from the isolated total RNA by reverse transcription followed by second strand synthesis using the Amino Allyl MessageAmp II aRNA Amplification kit (Ambion, Austin, TX) with oligo(dT) primer that contain a T7 RNA polymerase promoter sequence, according to the manufacturer’s specification. In vitro transcription was then performed with the purified cDNA and amino allyl UTP was incorporated during transcription to produce amino allyl modified aRNA that was subsequently coupled to cy5 dye label.
|
| |
|
| Hybridization protocol |
The HumanOneArray was hybridized using 2X Gex Hybridization Buffer (Agilent) for 17 h at 65oC; then washed in Gene Expression Wash Buffer 1 and 2 (Agilent) per manufacturer's instruction.
|
| Scan protocol |
The arrays were scanned with GenePix 4000B array scanner, images of the hybridized arrays were processed BioDiscovery's ImaGene and signal intensities quantified.
|
| Description |
none
|
| Data processing |
The raw data were re-scaled to account for the differences in individual hybridization intensities. We employed a rank-invariant scaling method to select genes (Tseng et al., 2001), which were then used for fitting of a non-linear normalization curve.
|
| |
|
| Submission date |
Dec 06, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Khew-Voon Chin |
| E-mail(s) |
khew-voon.chin@utoledo.edu
|
| Phone |
419-383-3502
|
| Fax |
419-383-4473
|
| Organization name |
The University of Toledo College of Medicine
|
| Department |
Medicine
|
| Street address |
3000 Arlington Avenue
|
| City |
Toledo |
| State/province |
OH |
| ZIP/Postal code |
43614 |
| Country |
USA |
| |
|
| Platform ID |
GPL6254 |
| Series (1) |
| GSE9806 |
A Time Course Analysis of the Effects of Marinobufagenin in Human Dermal Fibroblasts |
|
