|
| Status |
Public on Jan 29, 2011 |
| Title |
siPKCiz (+TGFbeta, 8 hours), biological replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
A549 cells transfected with siRNA directed at aPKC (PKCiota and PKCzeta) and treated with TGFbeta. RNA isolated 8 hours after TGFbeta treatment.
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: A549 human lung cancer cells from ATCC (ATCC number CCL-185)
transfection: siPKCiz
treatment: +TGFbeta
time: 8 hours
|
| Treatment protocol |
Control or aPKC silenced A549 cells were serum starved and then treated with 250 pM TGFbeta for 1 hour. Cells were then washed 3x with phosphate buffered saline and further incubated for an additional 7 or 23 hours in serum deprived media (0.2% FBS) prior to total RNA extraction.
|
| Growth protocol |
A549 Non small cell lung cancer cells were grown in F-12K media supplemented with 10% fetal bovine serum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed using the RNeasy Mini Kit (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated ds cDNA were prepared according to the Affymetrix Whole Transcript protocol from 200 ng total RNA (The Ambion WT Expression Kit 2009 and GeneChip WT Terminal Labeling and Hybridization User Manual, 2009-2010, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 5.5 ug of ss cDNA were hybridized for 16 hr at 45C on GeneChip Human Gene 1.0 ST Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G.
|
| Data processing |
Probe level data was generated using Affymetrix Command Console v1.1. Probes were summarized to gene level data in Partek Genomics Suite using the RMA algorithm.
|
| |
|
| Submission date |
Dec 21, 2010 |
| Last update date |
Jan 29, 2011 |
| Contact name |
John M Di Guglielmo |
| E-mail(s) |
[email protected]
|
| Phone |
519 661 2111 80042
|
| Organization name |
University of Western Ontario
|
| Department |
Phys/Pharm
|
| Lab |
MSB225
|
| Street address |
1151 Richmond
|
| City |
London |
| State/province |
ON |
| ZIP/Postal code |
N6A 5C1 |
| Country |
Canada |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE26241 |
Expression data from TGFbeta-treated control or aPKC silenced A549 cells |
|
