|
| Status |
Public on Oct 18, 2010 |
| Title |
Creb3L1 sorted 2 |
| Sample type |
RNA |
| |
|
| Source name |
HeLa cells; lipofectamine transfected with Creb3L1 T and GFP
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HeLa
treatment: lipofectamine transfected with Creb3L1 T and GFP
|
| Treatment protocol |
Cells were transfected with 1 ug of DNA using the Fugene 6.0 lipofectamine reagent (Roche) 20 hours prior to sorting. The amount of Creb3L1 T DNA was 0.8ug and DNA was 0.2ug.
|
| Growth protocol |
HeLa cells were grown at 37 C with 5%CO2 and humidity. Confluent cultures were split 24 hours prior to transfection. Following transfection, cells were grown in culture for 20 hours prior to removal for cell sorting.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using the Qiagen Rneasy kit.
|
| Label |
biotin
|
| Label protocol |
Amplification and labeling were performed using the Affymetrix GeneChip 3' IVT Express kit (P/N 901229) following the manufacturer's protocol.
|
| |
|
| Hybridization protocol |
Samples and controls were prepared using the Affymetrix GeneChip hybridization, wash and stain kit (P/N 900720). Arrays were prehybridized for 10 minutes prior to sample addition. Following sample addition, arrays were hybridized for 16 hours at 45 Celsius, and shaken at 60 rpm. Arrays were stained and washed using the Affymetrix GeneChip Fluidics Station 450 and FS450_002 fluidics script.
|
| Scan protocol |
Arrays were scanned with the Affymetrix GeneChip Scanner 3000 and raw analysis was performed with Affymetrix Command Console and Expression Console.
|
| Description |
Creb3L1 T/GFP expressing cells isolated by FACS using a FACSaria flow cytometer
|
| Data processing |
Data was normalized by RMA and statistical analysis was performed using Partek software. Further analysis was performed using Spotfire software.
|
| |
|
| Submission date |
Jul 30, 2010 |
| Last update date |
Oct 15, 2010 |
| Contact name |
Deborah J Andrew |
| E-mail(s) |
[email protected]
|
| Organization name |
Johns Hopkins University School of Medicine
|
| Department |
Cell Biology
|
| Street address |
725 N. Wolfe St., Hunterian G-1
|
| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21205 |
| Country |
USA |
| |
|
| Platform ID |
GPL6244 |
| Series (2) |
| GSE23334 |
Active Creb3L1 can upregulate secretory pathway genes in HeLa cells |
| GSE23349 |
The CrebA/Creb3-like transcription factors are major and direct regulators of secretory capacity |
|
