|
| Status |
Public on Feb 01, 2010 |
| Title |
Gata3_Cdx2_null_3 |
| Sample type |
RNA |
| |
|
| Source name |
Gata3 overexpression in Cdx2 null cells
|
| Organism |
Mus musculus |
| Characteristics |
Cdx2 null ES cells expressing human Gata3 T2ER fusion
|
| Treatment protocol |
To induce transgene activity, cells were treated with induction medium (TS medium + 1 µg/mL tamoxifen (Sigma)). For microarray analysis, ES cells were seeded at densities of 2 x 105 cells per gelatinized 35-mm well in ES medium, switched to induction medium the following day, and fed daily for the indicated time period.
|
| Growth protocol |
To create stably transformed lines, 1-3 x 107 ES cells were electroporated with 20-30 µg plasmid (pCAG-mCdx2ER-ires-puror (Niwa et al., 2005), pCAG-hCdx2ERT2-ires-puror, or pCAG-hGata3ERT2-ires-puror), and seeded on two 10 cm gelatinized plates. Cells were fed with plasmid selection medium (ES medium + 1.2 µg/mL puromycin) until colonies were visible by eye. Colonies were picked and individually expanded on gelatin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions. A second clean up step was done using the RNEasy kit from QIAGEN according to manufactures instructions
|
| Label |
biotin
|
| Label protocol |
as per facility protocols
|
| |
|
| Hybridization protocol |
as per facility protocols
|
| Scan protocol |
Gene Chips were scanned using the Hewlett-Packard Gene Array Scanner G2500A.
|
| Description |
Gata3 overexpression in Cdx2 null cells replicate 2
|
| Data processing |
The data were analyzed with Expression Console (Affymetrix) using the quartile normalization and PLIER algorithm
|
| |
|
| Submission date |
Sep 30, 2008 |
| Last update date |
Nov 23, 2009 |
| Contact name |
Brian Joseph Cox |
| E-mail(s) |
b.cox@utoronto.ca
|
| Organization name |
University of Toronto
|
| Department |
Physiology
|
| Lab |
Cox System Biology
|
| Street address |
1 King's College Circle, Rm 3360
|
| City |
Toronto |
| State/province |
Ontario |
| ZIP/Postal code |
M5S1A8 |
| Country |
Canada |
| |
|
| Platform ID |
GPL1261 |
| Series (2) |
| GSE12986 |
Expression of Cdx2 or Gata3 in R1 mouse embryonic stem cells |
| GSE12999 |
Gata3 acts alongside Cdx2 to promote trophoblast gene expression downstream of Tead4 during mouse development |
|
