|
| Status |
Public on Oct 28, 2009 |
| Title |
mouse natural helper cell 2 |
| Sample type |
RNA |
| |
|
| Source name |
unstimulated natural helper cells
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL6
tissue: Mesentery
|
| Treatment protocol |
Mesenteries separated from intestine and mesenteric lymph nodes were cut into small fragments with scissors and digested in Dulbecco's modified Eagle's Medium containing 2 mg/ml collagenase type I and 4% BSA. The supernatant was aspirated off after centrifugation to remove adipocytes. Finally, cells were suspended in Hank’s Balanced Salt Solution containing 10% FCS after filtration through 32 µm nylon mesh.
|
| Growth protocol |
Thy-1+CD4- LTi cells were prepared from fetal liver cells as described previously (J. Immunol. 167, 2511-2521, 2001). c-Kit+α4β7+IL-7Rα+ fetal liver cells from day 13 embryos were cultured on TSt4 cells for 17 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from 5 x 105 FALC c-Kit+Sca-1+ cells, LTi cells and DN2 cells after direct sorting into a vial containing ISOGEN LS (Nippon Gene, Tokyo, Japan). Total RNA of LTi cells was also extracted in ISOGEN LS. Total RNA was further purified using an RNeasy Micro Kit (QIAGEN, Tokyo, Japan) and amplified by Two-Cycle Target Labeling method (Affymetrix, Santa Clara, CA). Microarray processing was done by the Central Research Laboratory, Keio University School of Medicine.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 1 ug total RNA (Expression Analysis Technical Manual, ver. 6, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Mouse Genome Array 430 2.0. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 7G.
|
| Description |
mouse natural helper cell, mLSK2
|
| Data processing |
The data were analyzed with GeneChip Operation Software using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
Oct 27, 2009 |
| Last update date |
Aug 28, 2018 |
| Contact name |
kazuyo moro |
| E-mail(s) |
[email protected]
|
| Phone |
+81-3-5363-3769
|
| Organization name |
Keio University school of medicine
|
| Department |
Microbiology and Immunology
|
| Lab |
Koyasu
|
| Street address |
35
|
| City |
Sinjuku-ku Shinanomachi |
| State/province |
Tokyo |
| ZIP/Postal code |
160-8582 |
| Country |
Japan |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE18752 |
Gene expression profile from mouse natural helper cell |
|
| Relations |
| Reanalyzed by |
GSE45704 |
| Reanalyzed by |
GSE119085 |
