Gene expression in CD8α+ SPDCs and CD8α- SPDCs

Microarray analysis of CD8α+ SPDCs and CD8α- SPDCs from C57BL/6 mice; to elucidate genes specifically expressed in CD8α+ SPDCs CD8α+ DCs and CD8α- DCs were isolated from the spleen of C57BL/6 mice. Total RNA was extracted with an RNeasy kit (Qiagen), and purified using an Oligotex mRNA Kit (Pharmacia). Fragmented and biotin-labeled cDNA was synthesized from 100 ng purified mRNA using the Ovation Biotin System (Nugen), according to the manufacturer’s protocol. cDNA was hybridized to Affymetrix Murine Genome 430 2.0 microarray chips (Affymetrix) according to the manufacturer’s instructions. Hybridized chips were stained, washed and scanned using a GeneArray Scanner (Affymetrix). Data analysis was performed with Microarray Suite software (Version 5.0, Affymetrix) and GeneSpring software (Silicon Genetics).