|
| Status |
Public on Jun 18, 2010 |
| Title |
STAT4-deficient T cells, cultured 7 days in Th1 condition |
| Sample type |
RNA |
| |
|
| Source name |
primary CD4+ T cells from spleen and lymph nodes
|
| Organism |
Mus musculus |
| Characteristics |
cell: CD4+ T cell
genotype: C57BL6-STAT4<tm1> (STAT4KO)
|
| Treatment protocol |
After 7 day culture in vitro, cells were restimulated for 2hr with plate-bound anti-CD3, anti-CD28 (10 mg each) and IL-12 (20 ng/ml) for Th1 condition or IL-4 (10 ng/ml) for Th2 condition.
|
| Growth protocol |
Control and STAT-deficient naïve CD4+CD44-CD62L+ T cells were isolated and sorted on the flow cytometer. Cells were cultured for 72h with platebound anti-CD3, anti-CD28 (10 mg/ml each), IL-12 (20 ng/ml) and anti-IL-4 (10 mg/ml) for 3 days, followed by IL-2 (50 U/ml) and IL-12 (10 ng/ml) for 4 days (Th1 conditions), or anti-CD3, anti-CD28 (10 mg/ml each), IL-4 (10 ng/ml) and anti-IFN-g (10 mg/ml) for 3 days, followed by IL-2 (50 U/ml) and IL-4 (10 ng/ml) for 4 days (Th2 conditions).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the MirVana RNA Isolation kit from Ambion.
|
| Label |
biotin
|
| Label protocol |
RNA was biotin-labeled using the MessageAmpII-Biotin Enhanced Kit from Ambion
|
| |
|
| Hybridization protocol |
aRNA was hybridized to GeneChip Mouse Genome 430 2.0 Arrays from Affymetrix according the manufacturers protocol.
|
| Scan protocol |
Expression values were determined using GeneChip Operating Software (GCOS) v1.1.1 software.
|
| Description |
Gene expression data from STAT4-deficient CD4+ T cells cultured for 7 days in Th1 condition
|
| Data processing |
Data processing were done using Affymetrix Power Tool (APT) v. 1.10.2, quantile normalization RMA
|
| |
|
| Submission date |
Jun 01, 2010 |
| Last update date |
Jun 18, 2010 |
| Contact name |
Golnaz Vahedi |
| Organization name |
National Institutes of Health
|
| Department |
NIAMS
|
| Lab |
Lymphocyte Cell Biology Section
|
| Street address |
9000 Rockville Pike Bldg 10 Rm 13C101A
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (2) |
| GSE22081 |
Discrete Roles of STAT4 and STAT6 Transcription Factors in Tuning Epigenetic Modifications and Transcription during Helper T Cell Differentiation (gene expression) |
| GSE22105 |
Discrete Roles of STAT4 and STAT6 Transcription Factors in Tuning Epigenetic Modifications and Transcription during Helper T Cell Differentiation |
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