normalization data transformation protocol
Raw data were normalized by MAS 5.0 algorithm,GCOS1.4. ID_REF = VALUE = mas5 ABS_CALL = indicating whether the transcript was present (P), absent (A), or marginal (M) DETECTION P-VALUE =
array scanning protocol
GeneChips were scanned using the GeneChip® Scanner 3000.
Following fragmentation, cRNA were hybridized for 16 hr at 45C on GeneChip Mouse430 2.0 Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
cRNA were labeled by using One-Cycle Target Labeling and Control Reagents, Affymetrix, P/N 900493,and then purified by using QIAGEN RNeasy Total RNA Isolation kit.
sample treatment protocol
CD4+Foxp3+ Treg cells were sorted from the spleen of naive B6 DNR mice by FACS Aria.
nucleic acid extraction protocol
Total RNA was extracted from each sample using Trizol Plus (TAKARA) according to the manufacturer's protocol. RNA was then further purified using RNeasy Kit (Qiagen).