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Sample GSM1869261 Query DataSets for GSM1869261
Status Public on Nov 01, 2015
Title A.E7-stim for 5day-U74C2v
Sample type RNA
Source name A.E7 T cells stimulated for 16hrs and rested for 5days
Organism Mus musculus
Characteristics tissue: T cell clone
genotype: murine B10.A origin
treatment: anti-TCR antibody
time point: 5 days
Treatment protocol 5x10^6 cells were added to each well of the 6-well plate pre-coated with anti-TCR antbody. After 0, 2, 4 and 6hrs at 37C, 5% CO2, cells were harvested by scraping and frozen at -70C until RNA extraction. For the maintenance phase sample, cells were stimulated for 16hrs, washed three times, and incubated in fresh complete media at 37C, 5% CO2 without further stimulation for 5days before cell harvest and RNA extraction.
Growth protocol Cells were stimulated with whole pigeon cytochrome c protein in the presence of irradiated (3000 R) B10.A splenocytes as a source of APC. After 48 h, the cells were expanded in IL-2 (10U/ml) containing complete medium and rested for at least 2 weeks before restimulation or experimental use.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted using RNeasy kit(Qiagen) according to the manufacturer's instructions.
Label biotin
Label protocol Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 9.5 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
Hybridization protocol Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip U74 A then on B and last on C Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400
Scan protocol GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
Description cRNAs from AE.7 cells stimulated for 16hrs and rested for 5days were hybridized on mouse genome U74C(version2) chip.
Data processing The data were analyzed with Microarray Suite version 4.0 (MAS 4.0) using Affymetrix default analysis settings and global scaling as normalization method. The target intensity of each array was arbitrarily set to 150.
Submission date Sep 04, 2015
Last update date Nov 01, 2015
Contact name Kyungho Choi
Organization name Seoul National University College of Medicine
Department Department of Biochemistry and Molecular Biology
Lab Lab of Molecular Immunology
Street address 103 Daehak-ro, Jongno-gu
City Seoul
ZIP/Postal code 110-799
Country South Korea
Platform ID GPL83
Series (1)
GSE72731 Kinetic analysis of T cell anergy induction

Data table header descriptions
VALUE MAS4.0 signal intensity

Data table
128520_at 3094.6
128523_at 137.2
128529_at -186.1
128550_at -154.5
128554_at -146.9
128577_s_at 79.4
128578_r_at 550.8
128579_r_at 135.7
128580_at 179.3
128582_at 103.6
128583_at 14.3
128590_at 80.6
128624_at -83
128643_r_at -51.6
128644_at 593.2
128645_at -43.7
128646_at 208.9
128647_at -47.2
128651_at -10.9
128652_at 112.8

Total number of rows: 11934

Table truncated, full table size 191 Kbytes.

Supplementary file Size Download File type/resource
GSM1869261_KHC-1-Anerkinetics-5day-1-U74C2v.CEL.gz 1.8 Mb (ftp)(http) CEL
Processed data included within Sample table

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