|
| Status |
Public on May 15, 2015 |
| Title |
ROCK1 siRNA 3 |
| Sample type |
RNA |
| |
|
| Source name |
Human CASMCs, ROCK1 siRNA, 48 h
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: muscle
cell type: coronary artery smooth muscle
cell line: CC-2583
|
| Treatment protocol |
Human coronary artery smooth muscle cells were transfected with siRNA targeting ROCK1 or ZIPK or with negative control siRNA that does not target any gene product. 48 h later, total RNA was isolated.
|
| Growth protocol |
Human coronary artery smooth muscle cells (CC-2583, Lonza) were cultured in smooth muscle growth medium (Lonza SmGM-2: CC-3182) containing 5% fetal bovine serum, growth factors (hEGF, insulin and hFGF-β), gentamicin and amphotericin-B at 37 oC in a humidifed incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from the cells using the RNeasy Micro kit (Qiagen) according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
250 ng of RNA in each sample with RNA integrity number > 9 was reverse transcribed, amplified and labeled with the Ambion WT Express kit (Ambion).
|
| |
|
| Hybridization protocol |
Labeled cRNA was hybridized to Human Gene 1.0 ST arrays (Affymetrix) at 45 oC for 16 h. The probe arrays were washed and stained on an Affymetrix GeneChip Fluidics-450 following the manufacturer's instructions.
|
| Scan protocol |
GeneChips were scanned using Affymetrix GeneChip Scanner 3000 7G System.
|
| Description |
Gene expression data from ROCK1-knockdown cells
CASMC exposed to ROCK1 siRNA
|
| Data processing |
Array data files were generated with GeneChip® Command Console® Software (AGCC) and statistical significance was determined using GeneSpringTM (Agilent Technologies). The Fold Change between treatments and control was based on p<0.05 from Student's unpaired t-test.
|
| |
|
| Submission date |
Apr 15, 2014 |
| Last update date |
May 15, 2015 |
| Contact name |
Michael Patrick Walsh |
| E-mail(s) |
walsh@ucalgary.ca
|
| Phone |
403-220-3021
|
| Organization name |
University of Calgary
|
| Department |
Biochemistry & Molecular Biology
|
| Street address |
3330 Hospital Drive NW
|
| City |
Calgary |
| State/province |
Alberta |
| ZIP/Postal code |
T3A 0M9 |
| Country |
Canada |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE56819 |
Effects of knockdown of ROCK1 and ZIPK on gene expression in cultured human vascular smooth muscle cells |
|
