|
| Status |
Public on Jul 05, 2006 |
| Title |
Lbeta T2_1hr Activin |
| Sample type |
RNA |
| |
|
| Source name |
Lbeta T2 cell, a mature pituitary cell line.
|
| Organism |
Mus musculus |
| Characteristics |
A cell line developed by utilizing LHß promoter for targeted expression of SV40 T-antigen in transgenic mice
Expresses GnRH receptor, LHß and FSHß subunits, and secretes FSH upon activin stimulation
|
| Treatment protocol |
Recombinant human activin A was produced in our laboratory and formulated in a buffer of 0.15M NaCl and 0.05M Tris (Ph 7.5). Subconfluent cells were seeded in cell culture flasks for 16 hours in phenol-free, serum-free F12:DMEM medium, and then treated with activin at 30ng/ml.
|
| Growth protocol |
Cells were plated in cell culture flasks coated with matrigen (BD Biosciences, Bedford, MA) and maintained in F12:DMEM (Invitrogen) supplemented with 5% fetal bovine serum (FBS, Hyclone), 0.45% glucose and 100U of penicillin, and 100ug of streptomycin at 37C in 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Rneasy Mini Kit (Qiagen, MD) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10 microg total RNA (Expression Analysis Technical Manual, 2003, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 microg of cRNA were hybridized for 16 hr at 45C on Affymetrix mouse genechip 430 2.0. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix Agilent Genearray Scanner 3000.
|
| Description |
Gene expression data from Lbeta T2 cells treated with human activin A.
|
| Data processing |
The data were analyzed with Affymetrix Genechip Operating System version 1.4 (GCOS 1.4) and dCHIP using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 100.
|
| |
|
| Submission date |
Jul 05, 2006 |
| Last update date |
Jul 05, 2006 |
| Contact name |
Ying Peng |
| E-mail(s) |
yingp@northwestern.edu
|
| Organization name |
Northwestern University
|
| Street address |
2205 Tech Drive
|
| City |
Evanston |
| State/province |
IL |
| ZIP/Postal code |
60208 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE5232 |
Expression data from Lbeta T2 cells upon activin treatment |
|
