|
| Status |
Public on Apr 04, 2013 |
| Title |
8 = N16_KO_96h |
| Sample type |
RNA |
| |
|
| Source name |
hESCs_siREA_96h
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: primary cultured human endometrial stromal cells (hESCs)
transfected with: REA-specific siRNA for 24 h
treated with: [E2+P4+cAMP] hormone cocktail for 96hr
|
| Treatment protocol |
Independent hESC lines were established from individual biopsies of proliferative stage endometria, each cell line being from a different woman. These cultures were transfected with control (GL3 luciferase) or REA-specific siRNA for 24 h. siRNAs were then removed and cells were then treated for 24 h or 96 h in the presence of the hormone cocktail containing 10 nM E2, 1 µM progesterone and 0.5 µM 8Br-cAMP (E+P+cAMP). Cells were harvested and total RNA was prepared with TRIzol reagent. RNA integrity was verified using an Agilent 2100 Bioanalyser.
|
| Growth protocol |
Human endometrial stromal cells were maintained in DMEM/F-12 medium containing 5% charcoal stripped fetal bovine serum
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on Affymetrix [HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner 3000 7G Plus.
|
| Description |
4414_E
|
| Data processing |
The data were analyzed with GCOS2.1 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 |
Nov 19, 2012 |
| Last update date |
Apr 04, 2013 |
| Contact name |
Yuechao Zhao |
| Organization name |
University of Illinois
|
| Street address |
407 S. Goodwin Ave
|
| City |
Urbana |
| State/province |
IL |
| ZIP/Postal code |
61801 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE42376 |
The coregulator, repressor of Estrogen Receptor Activity (REA), is a crucial regulator of the timing and magnitude of uterine decidualization. |
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