|
| Status |
Public on Mar 16, 2006 |
| Title |
HVSMC_ Thapsigargin_treated_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
human cerebral artery
|
| Organism |
Homo sapiens |
| Characteristics |
Gender: Male
Age: 45 years
Tissue: human cerebral artery
|
| Biomaterial provider |
Human cerebral vascular smooth muscle cells (hcVSMCs) (passages 2-4) were obtained from human cerebral artery explants (IRB# CHRMS 01-195; informed consent) and maintained in SMGM2 (Clontech, Palo Alto, CA
|
| Treatment protocol |
Early passage hcVSMCs (passage 2-4) were obtained from human cerebral artery explants. Briefly, 2 mm slices of human pial artery (IRB# CHRMS 01-195; informed consent) were applied to scored 60 mm culture dishes, cultured in SMGM2 media (Cambrex Bio Science, Hopkinton, MA), and passaged with trypsin/EDTA. For hypoxia exposure, cells were incubated at 2% O2 by N2 injection into a humidified CO2 incubator (Forma, Marietta, OH). NOC-18, Cu/Zn superoxide dismutase (SOD), catalase, xanthine, and xanthine oxidase were obtained from Calbiochem (San Diego, CA). S-nitroso-L-glutathione (GSNO) was obtained from Alexis
|
| Growth protocol |
hcVSMCs and rat VSMCs (passages 2-4) were grown to approximately 60% confluence and serum starved in DMEM containing 0.1% FBS 24-48 hr prior to treatment
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from treated hc VSMCs using TriZol reagent and chloroform. RNA was precipitated using isoproponol and glycoblue, washed with 75% ethanol, dissolved in RNAse free water and quantified using the NanoDrop® spectrophotometer.
|
| Label |
biotin-labeled
|
| Label protocol |
Double-stranded cDNA was then synthesized followed by an in vitro transcription reaction and fragmentation reaction to produce biotin-labeled cRNA fragments that were hybridized to the probe array at 45°C for 16 hr
|
| |
|
| Hybridization protocol |
biotin-labeled cRNA fragments that were hybridized to the probe array at 45°C for 16 hr
|
| Scan protocol |
The probe arrays were scanned (Hewlett-Packard GeneArray Scanner, Agilent Technologies, Inc.) to quantify the fluorescence intensity associated with each oligonucleotide probe.
|
| Description |
each of three experiments cell cultures were split three ways; one of the resulting samples was left untreated (C), another was treated with thapsigargin (TG), and the third was treated with elevated K+ (K)
|
| Data processing |
RMA
|
| |
|
| Submission date |
Jul 06, 2005 |
| Last update date |
Mar 16, 2006 |
| Contact name |
Anjanette D Watson |
| E-mail(s) |
[email protected]
|
| Phone |
8026568612
|
| Organization name |
University of Vermont
|
| Department |
Bioinformatics Core
|
| Street address |
120A Marsh Life Science
|
| City |
Burlington |
| State/province |
VT |
| ZIP/Postal code |
05405 |
| Country |
USA |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE2883 |
Ca2+-dependent Transcription Patterns in Human Cerebrovascular Smooth Muscle |
|
