|
| Status |
Public on Dec 22, 2020 |
| Title |
WT_Resting_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Splenic B cells from B1-8hi:Sub1fl/fl
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Splenic B cells
treatment: Naïve
genotype/variation: B1-8hi:Sub1fl/fl
|
| Treatment protocol |
Splenic B cells were cultured in RPMI-1640 medium supplemented with 100 U/mL IL-2, 5 ng/mL IL-4, 1.5 ng/mL IL-5, 0.2 ng/mL CD40 ligand, 0.01 ng/mL NP-ficoll, 10% FBS 10mM HEPES, 1 mM Na-Pyruvate, 0.1 mM non-essential amino acids and penicilli/streptmycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the RNeasy Plus Micro kit (QIAGEN) following the manufacturer's recommendations. RNA quantity and quality were measured with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Total RNA was amplified with an Amino Allyl MessageAmpII aRNA Amplification Kit (Ambion) and was labeled with indocarbocyanine through the use of a CyDye post-labeling Reactive Dye Pack (Amersham).
|
| |
|
| Hybridization protocol |
A sureprint G3 mouse GE microarray slide was used for hybridization according to the manufacturer's instructions(Agilent).
|
| Scan protocol |
Agilent scan protocol
|
| Description |
Gene expression without differentiation stimuli
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters (protocol GE1_107_Sep09 and Grid: 028005_D_F_20101029) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Feb 26, 2020 |
| Last update date |
Dec 22, 2020 |
| Contact name |
Kyoko Ochiai |
| E-mail(s) |
kochiai@med.tohoku.ac.jp
|
| Organization name |
Tohoku University School of Medicine
|
| Department |
Biochemistry
|
| Street address |
2-1, Seiryo-machi, Aobaku
|
| City |
Sendai |
| ZIP/Postal code |
980-8575 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13912 |
| Series (2) |
| GSE145952 |
Chromatin Protein PC4 Orchestrates B Cell Differentiation by Collaborating with IKAROS and IRF4 (Agilent microarrays) |
| GSE145953 |
Chromatin Protein PC4 Orchestrates B Cell Differentiation by Collaborating with IKAROS and IRF4 |
|
