|
| Status |
Public on Apr 16, 2011 |
| Title |
Non-phosphorylated FTY720 induces apoptosis of human microglia by activating SREBP2(#2) |
| Sample type |
RNA |
| |
|
| Source name |
HMO6 human microglia treated with FTY720 dissolved in DMSO for 2 hours
|
| Organism |
Homo sapiens |
| Characteristics |
cells: HMO6 microglia
tissue: Brain
|
| Biomaterial provider |
Seung U. Kim, Division of Neurology, Department of Medicine, University of British Columbia Hospital, University of British Columbia, Vancouver, British Columbia, Canada
|
| Treatment protocol |
HMO6 cells were exposed for 2 hours to 10 microM non-phosphorylated FTY720 (Calbiochem) or an equal amount of vehicle (DMSO).
|
| Growth protocol |
The cells were maintained in Dulbecco's Modified Eagle's medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 microgram/ml streptomycin (feeding medium).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from the cells by using the TRIZOL Plus RNA Purification kit (Invitrogen). The quality of total RNA was evaluated on Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
biotin
|
| Label protocol |
One hundred ng of total RNA was processed for cRNA synthesis, fragmentation, and terminal labeling with the GeneChip Whole Transcript Sense Target Labeling and Control Reagents (Affymetrix)
|
| |
|
| Hybridization protocol |
Hybridization was performed at 45℃ for 17 hours. The arrays were washed and stained in the GeneChip Fluidic Station 450 (Affymetrix) by using Hybridization, Wash and Stain kit (Affymetrix).
|
| Scan protocol |
The arrays were scanned by the GeneChip Scanner 3000 7G (Affymetrix).
|
| Description |
The HMO6 cell line was established by immortalizing cultured microglia isolated from human embryonic telencephalon tissues with a retroviral vector PASK1.2 encoding v-myc oncogene. HMO6 cells express the markers of the microglia/macrophage lineage cells, including CD11b, CD68, CD86, HLA-ABC, HLA-DR, and ricinus communis agglutinin lectin-1 (RCA), serving as a model of human microglia both in vitro and in vivo.
|
| Data processing |
The data expressed as CEL files were normalized by the robust multiarray average (RMA) method with the Expression Console software version 1.1 (Affymetrix).
|
| |
|
| Submission date |
Apr 15, 2011 |
| Last update date |
Apr 16, 2011 |
| Contact name |
Jun-ichi Satoh |
| E-mail(s) |
satoj@my-pharm.ac.jp
|
| Organization name |
Meiji Pharmaceutical University
|
| Department |
Bioinformatics
|
| Lab |
Molecular Neuropathology
|
| Street address |
2-522-1 Noshio, Kiyose
|
| City |
Tokyo |
| ZIP/Postal code |
204-8588 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE28642 |
Non-phosphorylated FTY720 induces apoptosis of human microglia by activating SREBP2 |
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