Genetic background: BDC2.5/NOD Age: 4-6 wks Number of mice per chip: 3-6 Tissue: Spleen and LN (cervical, mesenteric) Sort: B220-, CD11b-, CD8a-, CD69-, CD62Lhi, CD4+, CD25- Culture: Activation in vitro with anti-CD3/-CD28 beads + IL2 (20 U/ml) + TGFbeta (25ng/ml) for 24hrs, then spin infected with retrovirus (empty-GFP) and cultured for an additional 72hrs. Post-culture sort: GFP+ cells
Extracted molecule
total RNA
Extraction protocol
RNA isolated with Trizol reagent and amplified twice with the MessageAmp aRNA Kit (Ambion)
Label
Biotin
Label protocol
BioArray High Yield RNA Transcript Labeling T7 Kit (Enzo Life Sciences)
Hybridization protocol
Per Joslin Diabetes Center microarray core facility.
Scan protocol
Per Joslin Diabetes Center microarray core facility.
