|
| Status |
Public on Nov 14, 2016 |
| Title |
Bach2_KO_IL-2_(+) |
| Sample type |
RNA |
| |
|
| Source name |
Bach2 KO Th cells re-stimulated with immobilized anti-TCR-β mAb
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57BL/6J
genotype/variation: Bach2 KO
gender: female
tissue: spleen
cell type: Primary naïve CD4 T cells
|
| Growth protocol |
Bach2 WT and Bach2 KO (Bach2flox/flox × CD4-Cre TG) naïve CD4 T (CD44lowCD62LhighCD25negative) cells were prepared using a CD4+CD62L+ T cell isolation kit II (Miltenyi Biotec). Naïve CD4 T cells were stimulated with immobilized anti-TCR-β mAb (3 μg ml-1, H57-597; BioLegend) and an anti-CD28 mAb (1 μg ml-1, 37.5; BioLegend) for two days under the conditions indicated. Next, the cells were transferred to a new plate and cultured for an additional three days in the presence of cytokines. Th cells were re-stimulated with immobilized anti-TCR-β mAb for four hours in the absence of exogenous IL-2. The cytokine conditions were as follows: IL-2 conditions (Th), IL-2 (2.5 ng ml-1)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated with TRIZOL regent (invitrogen)
|
| Label |
Cy3
|
| Label protocol |
500 ng of total RNA was amplified and labeled using the One-Color Agilent Quick Amp Labeling kit
|
| |
|
| Hybridization protocol |
Hybridization and washing were done according to Agilent One-Color Gene Expression protocol
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent Microarray Scanner (G2565CA) using one color scan setting
|
| Data processing |
Raw signal intensities were extracted with Agilent Feature Extraction Software v10.7 using default parameters (protocol GE1-107_Sep09). The dataset was normalized using the 75 percentile median shift method.
|
| |
|
| Submission date |
Jan 19, 2015 |
| Last update date |
Nov 14, 2016 |
| Contact name |
Makoto Kuwahara |
| E-mail(s) |
mkuwa@m.ehime-u.ac.jp
|
| Organization name |
Graduate School of Medicine, Ehime University
|
| Department |
Department of Immunology
|
| Lab |
Immunology
|
| Street address |
Shitsukawa
|
| City |
Toon-shi |
| State/province |
Ehime |
| ZIP/Postal code |
791-0295 |
| Country |
Japan |
| |
|
| Platform ID |
GPL7202 |
| Series (2) |
| GSE65083 |
Bach2 inhibits Th2-type immune responses by regulating glutamine metabolism [expression array] |
| GSE65084 |
Bach2-Batf interactions control Th2-type immune responses by regulating the IL-4 amplification loop |
|
