|
| Status |
Public on Feb 09, 2011 |
| Title |
STAT5-deficient T cells, cultured 3 days in Th17 condition (expression) |
| Sample type |
RNA |
| |
|
| Source name |
primary naïve CD4+ T cells from spleen and lymph nodes
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
genotype/variation: CD4-Cre stat5 fl/fl OT2 (STAT5 wild type)
cell type: primary naïve CD4+ T cells from spleen and lymph nodes
|
| Treatment protocol |
After 3 day culture in vitro, cells were harvested and lysised with 300 ul lysis buffer from the MirVana RNA Isolation kit.
|
| Growth protocol |
Control and STAT-deficient naïve CD4+CD44-CD62L+ T cells were isolated and sorted on the flow cytometer. Cells from STAT3 WT and STAT3 KO were cultured for 72h with platebound anti-CD3, anti-CD28 (10 mg/ml each), IL-6 (20 ng/ml) TGF-beta (2 ng/ml) , anti-IL-2 (10 ug/ml) and anti-IL-4 (10 mg/ml) for 3 days. Cells from STAT5 WT and STAT5 KO were cultured for 72h with platebound anti-CD3, anti-CD28 (10 mg/ml each), IL-6 (20 ng/ml) TGF-beta (2 ng/ml) , IL-2 (100IU/ml) and anti-IL-4 (10 mg/ml) for 3 days.
|
| 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 |
According the manufacturers protocol
|
| Data processing |
Affymetrix Power Tools using apt-probeset-summarize & rma-sketch.
|
| |
|
| Submission date |
Jan 11, 2011 |
| Last update date |
Feb 09, 2011 |
| 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) |
| GSE26551 |
Roles of STAT3 and STAT5 in regulation of gene expression under Th17 differentiation |
| GSE26553 |
Opposing regulation of the Il17 locus through direct, reciprocal actions of STAT3 and STAT5 |
|
