Gene expression induced by flagellin stimulation in IECs. Mus musculus

Microarray analysis of IECs from Tlr5+/+ and Tlr5–/– mice stimulated with either medium alone or flagellin (1 µg/ml); to elucidate TLR5-mediated immune responses in IECs. Keywords: ordered Overall design: IECs from Tlr5+/+ and Tlr5–/– mice were treated with or without flagellin (1 ug/ml) for 4 h. 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).