Identification of microRNAs involved in NOD-dependent induction of pro-inflammatory genes in pulmonary endothelial cells

Primary lung endothelial cells were isolated from wildtype mice and stimulated with the NOD1 ligand TD or NOD2 ligand MDP. Unstimulated samples were used as controls. RNA was extracted from these samples, reverse transcribed and microRNA expression was analyzed using TLDA. 350 ng RNA per sample were then reverse transcribed using TaqMan MicroRNA Reverse Transcription Kit and Megaplex RT Primers for rodent Pool A. Reverse transcribed RNA was then loaded on the Taqman Low Density Array (TLDA SetA, v.2.0 rodent, Life Technologies) Cards and run according to manufacturer´s recommendations. microRNA profiles were used to identify NOD-regulated endothelial cell microRNAs and in silico target genes