|
| Status |
Public on May 02, 2012 |
| Title |
Mouse bone marrow-derived mast cells treated with IL3+SCF, rep 4 |
| Sample type |
RNA |
| |
|
| Source name |
mouse bone marrow-derived mast cells treated with IL3+SCF
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
cell type: bone marrow-derived mast cells
treatment: interleukin 3 and stem cell factor
|
| Treatment protocol |
Essentially, the cells were cultured for 4-6 weeks in media containing mouse recombinant IL-3 (30 ng/ml) (Peprotech, Rocky Hill, NJ) and mouse recombinant SCF (unless otherwise indicated: 100 ng/ml) (Peprotech). The cells were maintained at 37 ˚C in a humidified incubator gassed with 95% air and 5% CO2. The purity of the cultures, as assessed by toluidine blue staining and FceRIa and KIT expression, was >99%.
|
| Growth protocol |
Bone marrow-derived mast cells (BMMCs) were developed from bone marrow obtained from the femurs of C57BL/6 mice (The Jackson Laboratory, Bar Harbor, ME) as described (Kuehn, H. S., M. A. Beaven, H. T. Ma, M. S. Kim, D. D. Metcalfe, and A. M. Gilfillan. 2008. Synergistic activation of phospholipases Cγ and Cβ: a novel mechanism for PI3K-independent enhancement of FcεRI-induced mast cell mediator release. Cell Signal 20:625-636 (PMID 18207701)).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNAs were extracted using an RNeasy 96 kit (Qiagen, Valencia, CA) with each sample treated with 27 units DNase I during the extraction process.
|
| Label |
biotin
|
| Label protocol |
Affymetrix GeneChip targets were made by amplification of 50 ng RNA using the WT-Ovation RNA Amplification System (Nugen Inc., San Carlos, CA).
|
| |
|
| Hybridization protocol |
Hybridization and fluidics were performed according to standard Affymetrix protocols (http://www.affymetrix.com).
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 7GPlus according to standard Affymetrix protocols.
|
| Description |
Gene expression data from mouse bone marrow-derived mast cells treated with IL3+SCF, rep 4 of 6.
|
| Data processing |
The data were analyzed with GCOS 1.4 and EC 1.1 using Affymetrix default settings except the trimmed mean target intensity of each array was arbitrarily set to 1250.
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| |
|
| Submission date |
Jan 25, 2012 |
| Last update date |
Jul 16, 2012 |
| Contact name |
Dan Sturdevant |
| E-mail(s) |
[email protected]
|
| Phone |
4063639248
|
| Organization name |
NIH
|
| Department |
NIAID
|
| Lab |
RTS
|
| Street address |
903 S 4th street
|
| City |
Hamilton |
| State/province |
MT |
| ZIP/Postal code |
59840 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE35332 |
Stem cell factor programs the mast cell activation phenotype |
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