|
| Status |
Public on Nov 07, 2016 |
| Title |
MH7A IL-1β-stimulated ex2 |
| Sample type |
RNA |
| |
|
| Source name |
IL-1β-stimulated MH7A
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MH7A
cell type: synovial cell line
treatment group: IL-1β-stimulated MH7A
|
| Treatment protocol |
Cells (60-70% confluent) in 6-well plates were incubated in the presence or absence of 5 mM GlcN (Koyo Chemical, Japan) foｒ 2.5 hours, and then stimulated with 15 pg/ml IL-1β for 13h. In some experiments, cells were preincubated with 5 mM alloxan (O-GlcNAc transferase inhibitor) for 30 min, and then incubated with 2 mM alloxan and 5mM GlcN for 2.5 h, followed by stimulation with IL-1β. Thereafter, the cells were harvested, and total RNA was purified.
|
| Growth protocol |
Human synovial MH7A cells were maintained in RPMI-1640 medium containing 10% fetal bovine serum, 100 units/ml penicillin and 100 µg/ml. streptomycin at 37°C in 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was purified using a QIA shredder (QIAGEN) and an RNeasy plus mini kit (QIAGEN) according to the manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared using WT Terminal Labeling Kit (Affymetrix) according to the manufacturer’s instructions from100 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 5.5 µg of cDNA were hybridized for 17 hr at 45℃ on GeneChip Human Gene 1.0 ST Array. GeneChips were washed and stained in the Affymetrix GeneChip Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G.
|
| Description |
MH7A Control 2-2
Gene expression data from IL-1β-stimulated MH7A
|
| Data processing |
The data were analyzed with Affymetrix Expression Console using Robust Multi-array Average (RMA) summarization method.
affymetrix-algorithm-name = rma-gene-default
affymetrix-algorithm-version = 1.0
program-name = Expression Console
program-version = 1.1.2800.19935
|
| |
|
| Submission date |
Sep 01, 2015 |
| Last update date |
Nov 07, 2016 |
| Contact name |
AKIMASA SOMEYA |
| E-mail(s) |
someya@juntendo.ac.jp
|
| Phone |
+81-3-5802-1033
|
| Organization name |
Juntendo University
|
| Department |
Department of Host Defense and Biochemical Research
|
| Street address |
2-1-1 Hongo
|
| City |
Bunkyo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
113-8421 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE72575 |
Expression data from glucosamine-treated human synovial MH7A cells |
|
