|
| Status |
Public on Jun 06, 2017 |
| Title |
T-helper cell population Th2, miRNA, biological replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Th2 cells derived from CD4 positive spleen lymphocytes
|
| Organism |
Mus musculus |
| Characteristics |
strain: Balb/c
genotype/variation: WT
age: 10-14 weeks
gender: female
t-helper cell source: CD4 positive spleen lymphocytes
t-helper populations: Th2 cells
|
| Treatment protocol |
Cells populations were treated as described in the paper. Briefly, isolation of CD4+ lymphocytes from spleens of Balb/c mice were conducted with the mouse CD4+ Isolation Kit II from Miltenyi Biotechnology. 2*105 CD4+ cells were seeded in one well of a 96- well plate. All cells were cultured with RPMI1640 media containing 10% FCS, 1% penicillin/streptomycin and antibodies against CD3 (4µg/ml) and CD28 (30ng/ml) (BioLegend). Th17 cells were additionally cultured with IL-6(20ng/ml), TGFβ (5ng/ml), IL-23 (10ng/ml) (R&D Systems) and an antibody against IFNγ (10µg/ml) (BioLegend). Th2 cells were differentiated with IL-4 (100ng/ml) (R&D Systems) and an antibody against IFNγ (10µg/ml) (BioLegend). Th0 cells serve as a control aand were cultured with solely anti-CD3 and anti-CD28 antibodies. The primary Th cells were cultivated at 37°C and 5% CO2 in a humidified incubator. After 72h the media was exchanged by fresh differentiation media.
|
| Growth protocol |
Animals per housed under spf conditions
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRNeasy (Qiagen) including DNase treatment
|
| Label |
biotin
|
| Label protocol |
FlashTag Biotin HSR kit (Genisphere)
|
| |
|
| Hybridization protocol |
According to the Affymetrix expression protocol (HWS kit)
|
| Scan protocol |
According to the Affymetrix expression protocol
|
| Description |
Th2_miRNA_4
miRNA expression data from T-helper cell population Th2
|
| Data processing |
RMA probeset summary with standard settings (RMA+DABG) of the Affymetrix Expression Console software (v.1.3.0.187)
|
| |
|
| Submission date |
Feb 13, 2014 |
| Last update date |
Jun 06, 2017 |
| Contact name |
Johannes Beckers |
| E-mail(s) |
johannes.beckers@helmholtz-munich.de
|
| Organization name |
Helmholtz Zentrum Muenchen
|
| Department |
Institute of Experimental Genetics
|
| Street address |
Ingolstaedter Landstr. 1
|
| City |
Neuherberg |
| ZIP/Postal code |
85764 |
| Country |
Germany |
| |
|
| Platform ID |
GPL16384 |
| Series (2) |
| GSE55012 |
microRNA cluster 106a~363 is involved in T helper 17 cell differentiation |
| GSE55013 |
Profiling of in vitro differentiated activated T helper cells |
|
