|
| Status |
Public on Jan 24, 2013 |
| Title |
Tot2_MSCV-IRF8-rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Tot2 cells, MSCV-IRF8 transuced, Day4, replicate 2
|
| Organism |
Mus musculus |
| Characteristics |
cell line: Myeloid progenitor cell line Tot2 (derived from Irf8 knockout mouse)
treatment period: 4 days
culture condition: MSCV-IRF8 transduced
|
| Treatment protocol |
Retroviral preparation and transduction with control MSCV-puro, MSCV-IRF8FLAG or MSCV-KLF4FLAG were performed as described previously (Tamura et al. Blood 106: 1938, 2005). Cells were transduced by spinoculation in a retroviral supernatant supplemented with cytokines and 4 μg/ml polybrene. Transduced cells were selected with 2 μg/ml puromycin 48 hr after spinoculation. At 4 days after the transduction, the cells were harvested for RNA isolation.
|
| Growth protocol |
Myeloid progenitor cell line Tot2 cells derived from Irf8 knockout mouse were cultured in RPMI1640 supplemented with 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 20% fetal bovine serum and 5.0 ng/ml granulocyte-macrophage colony stimulating factor.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells using RNAiso Plus reagent (Takara) and further purified using RNeasy Mini Kit (Qiagen) according to the manufacturers’ instructions.
|
| Label |
Cy3
|
| Label protocol |
Total RNA was labeled using the Agilent Low Input Quick Amp Labeling Kit according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Slides were hybridized according to the manufacturer's protocol (One-Color Microarray-Based Gene Expression Analysis Protocol, Product # G4140-90040, Version 6.5, May 2010).
|
| Scan protocol |
Slides were scanned with an Agilent DNA Microarray Scanner (model G2505-60502) at 100% in Green channel, with a scan resolution of 3um.
|
| Data processing |
Data are extracted with Agilent Feature Extraction Software. Each measurement was divided by the 75th percentile of all measurements in that sample for normalization. Microarray data in two replications was further processed with analyzed by NIA Array Analysis software (http://lgsun.grc.nia.nih.gov/ANOVA/).
|
| |
|
| Submission date |
Jun 19, 2012 |
| Last update date |
Jan 24, 2013 |
| Contact name |
Tomohiko Tamura |
| E-mail(s) |
tamurat@yokohama-cu.ac.jp
|
| Organization name |
Yokohama City University
|
| Department |
Department of Immunology
|
| Street address |
3-9 Fukuura, Kanazawa-ku
|
| City |
Yokohama |
| ZIP/Postal code |
236-0004 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10787 |
| Series (2) |
| GSE38810 |
Gene expression analyses in IRF8-induced monocyte differentiation |
| GSE38825 |
IRF8-induced monocyte differentiation |
|
