|
| Status |
Public on Oct 11, 2008 |
| Title |
Mouse PUER exon 0h-1 |
| Sample type |
RNA |
| |
|
| Source name |
PU-ER cells without expression of PU.1
|
| Organism |
Mus musculus |
| Characteristics |
PU.1 -/- deficient myeloid progenitors with PU-ER fusion protein
|
| Treatment protocol |
To activate the PU.1-ER fusion protein, PUER cells were centrifuged and resuspended in medium containing 100nM tamoxifen [4-hydroxytamoxifen (OHT)]
|
| Growth protocol |
PUER cells were cultivated in IMDM supplemented with penicillin/streptomycin (10,000 U/ml), glutamin (200 mM), ß-ME (50 M), mouse IL-3 (5 ng/ml, Biosource, Camarillo, CA, USA), puromycin (1 g/ml), and 10% FCS.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from the PUER cells was extracted according to the manufacter's instructions using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA)
|
| Label |
Biotin
|
| Label protocol |
Biotinylated sense-strand DNA for hybridization to Affymetrix exon arrays was prepared with the Affymetrix GeneChip whole transcript sense target labeling assay, using the standard 1,5 μg total RNA method according to the manufacturer’s protocols (Affymetrix, GeneChip WT terminal labelling kit 900671). Ribosomal RNA reduction was performed using the RiboMinus Human/Mouse Transcriptome Isolation Kit (Invitrogen, K1550-01).
|
| |
|
| Hybridization protocol |
The labeling assay yielded labeled single-stranded DNA that was fragmented and hybridized to Affymetrix Mouse Exon 1.0 ST Arrays (exon arrays) according to manufacturer’s protocols at the KFB Regensburg
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 Blank7G (Affymetrix).
|
| Description |
Gene expression data from PU-ER cells without expression of PU.1.
|
| Data processing |
The data were analyzed with Affymetrix Expression console and Genomatix ChipInspector using Affymetrix default analysis settings and global scaling as normalization method.
MoEx-1_0-st-v1.r2.dt1.mm8.core.mps
|
| |
|
| Submission date |
Oct 08, 2008 |
| Last update date |
Oct 10, 2008 |
| Contact name |
Thomas Langmann |
| E-mail(s) |
thomas.langmann@klinik.uni-regensburg.de
|
| Phone |
+49-941-944-5423
|
| Fax |
+49-941-944-5402
|
| Organization name |
Institute of Human Genetics
|
| Department |
University of Regensburg
|
| Street address |
Franz-Josef-Strauss-Allee 11
|
| City |
Regensburg |
| ZIP/Postal code |
93042 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6096 |
| Series (1) |
| GSE13125 |
Identification of PU.1 target genes by expression profiling of PUER cells |
|
