|
| Status |
Public on Aug 03, 2012 |
| Title |
CD41hi, A2lox.Meis2 and OP9-GFP co-culture, D8, treated with dox |
| Sample type |
RNA |
| |
|
| Source name |
GFP-CD41hi cells purified on day 8 from A2lox.Meis2 and OP9-GFP co-culture treated with doxycycline for 48 hours
|
| Organism |
Mus musculus |
| Characteristics |
cell type: dox-inducible transgenic A2lox.Meis2 mouse embryonic stem cells
background strain: 129/Ola
day of differentiation: 8
treatment: doxycycline on day 6
population: GFP-CD41hi
|
| Treatment protocol |
ES cell and OP9-GFP cocultures were treated with (+) or without (-) doxycycline (dox) at a final concentration of 500 ng/ml on day 6. On day 7, ES cell and OP9-GFP cocultures were purified into GFP-CD41- and GFP-CD41+ populations by cell sorting on the FACS Aria II. On day 8, ES cell and OP9-GFP cocultures were purified into GFP-CD41-, GFP-CD41int and GFP-CD41hi populations by cell sorting on the FACS Aria II.
|
| Growth protocol |
A2lox.Meis2 ES cells were differentiated as embryoid bodies (EBs) for 6 days. On day 6, EBs were dissociated and plated at a density of 100,000 cells/ml on irradiated OP9-GFP cell monolayers with cytokines (100 ng/ml SCF, 40 ng/ml TPO, 40 ng/ml VEGF, 5% Flt3-L conditioned media, 10 ng/ml IL-3 and 20 ng/ml IL-6).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA from sorted cell populations was extracted using Qiagen's RNeasy kits according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Amplified cDNA was prepared using NuGEN Ovation PicoSL WTA System from 50 ng total RNA. The cDNA product was then fragmented and labeled according to a validated NuGEN protocol.
|
| |
|
| Hybridization protocol |
The Affymetrix operating system (Command Console) was used to manage the washing and hybridization steps.
|
| Scan protocol |
The Affymetrix operating system (Command Console) was used for scanning. Affymetrix analysis software (Genotyping Console) was used to derive Quality Control metrics, create .CEL files and scaled expression data from the scanned expression array image files.
|
| Description |
Meis2+dox_D8_CD41hi
Expression data from GFP-CD41hi cells purified on day 8 from A2lox.Meis2 and OP9-GFP co-culture treated with doxycycline for 48 hours.
|
| Data processing |
The data was analyzed using DNASTAR ArrayStar 4 software. RMA was selected as the preprocessing method and Quantile was selected as the preprocessing parameters.
|
| |
|
| Submission date |
Dec 19, 2011 |
| Last update date |
Aug 03, 2012 |
| Contact name |
Mi Cai |
| E-mail(s) |
mcai@wustl.edu
|
| Phone |
314-362-2004
|
| Fax |
314-747-4888
|
| Organization name |
Washington University School of Medicine
|
| Department |
Immunology and Pathology
|
| Lab |
Ken Murphy
|
| Street address |
660 S. Euclid Ave.
|
| City |
St. Louis |
| State/province |
MO |
| ZIP/Postal code |
63110 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (2) |
| GSE34541 |
Identification of gene targets of Meis2 |
| GSE34583 |
Dual actions of Meis1 inhibit erythroid progenitor development and sustain general hematopoietic cell proliferation |
|
