|
| Status |
Public on May 10, 2011 |
| Title |
H20 HA 12h 4 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse forebrain primary cell cultue of astrocytes
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57 Bk6
age: 1.5 day
tissue: forebrain
cell type: primary cell culture astrocyte
treatment: HA1077
time: 12h
culture batch: 2
bead chip array sentrix code: 4421339033
|
| Treatment protocol |
Monolayers of astrocytes were treated with Fasudil (diluted in astrocytic medium pre-warmed at 37ºC) and the volume was maintained at 0.5 ml per well. Vehicle-treated cultures were run in parallel to serve as controls.
|
| Growth protocol |
Cells plated in 24-well plates (2.6 × 104 cells/cm2) and incubated in a humidified incubator at 36.5ºC with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted in one batch using the RNeasy Mini Kit and quality of the extracted RNA was determined using E-gene HAD-GT12 genetic analyser (John Morris Scientific, Melbourne, Australia).
|
| Label |
biotin
|
| Label protocol |
RNA was amplified using the Illumina® TotalPrep RNA Amplication Kit. Total RNA sample was reverse transcribed using T7 Oligo(dT) primer to generate first strand second strand cDNA. Biotinylated cRNA was generated from the double-stranded cDNA templates via in vitro transcription.
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
replicate 4
|
| Data processing |
Gene expression data was exported from Bead Studio using median shift normalization such that the lowest value is 1 and log2 transformed with the ‘Partek report plug-in’ (Partek® Inc., St. Louis, Missouri, U.S.A.) and analysed in Partek Genomics Suite. The resultant export file in project file format (.ppj) included log transformed signal intensity data and annotation. This file was then imported into Partek Genomics suite Version 6.3 (Partek® Inc., St. Louis, Missouri, U.S.A.). The normalised expression data was then exported from Partek, signal p values were not available for normalised expression values.
|
| |
|
| Submission date |
Dec 02, 2010 |
| Last update date |
May 10, 2011 |
| Contact name |
Victoria Mary Perreau |
| E-mail(s) |
vperreau@unimelb.edu.au
|
| Phone |
+61 3 8344 1835
|
| Organization name |
University of Melbourne
|
| Department |
Center for Neuroscience
|
| Street address |
Parkville
|
| City |
Melbourne |
| State/province |
Victoria |
| ZIP/Postal code |
3010 |
| Country |
Australia |
| |
|
| Platform ID |
GPL6885 |
| Series (1) |
| GSE25829 |
TRANSCRIPTOMIC PROFILING OF ASTROCYTES TREATED WITH THE RHO KINASE INHIBITOR FASUDIL REVEALS CYTOSKELETAL AND PRO-SURVIVAL RESPONSESTRANSFORMATION INDUCED BY THE RHO KINASE INHIBITOR FASUDIL. |
|
