Transcription profiling by array of human dendritic cells treated with rosiglitazone and/or AGN193109

Our data indicated that activation of the PPARg nuclear receptor induces a retinoid response in human dendritic cells. In order to assess the contribution of retinoid signaling to the PPARg response we decided to use a combination of pharmacological activators and inhibitors of these pathways. Cells were treated with the synthetic PPARg ligand rosiglitazone (RSG), or with RSG along with the RARa antagonist (AGN193109) to block RARa mediated gene expression, or the RARa specific agonists (AM580) alone. This design allows one to determine if retinoid signaling is a downstream event of PPARg activation and what portion of PPARg regulated genes are regulated via induced retinoid signaling. Experiment Overall Design: Monocytes were cultured for 5 days with 500 U/ml IL-4 and 800 U/ml GM-CSF, cytokine treatment was repeated at day 3. Ligands were added at the beginning of differentiation but AGN193109 (AGN) treatment was repeated at day 3. Cells were obtained from three healthy individuals (three biological replicates) and cells were treated with vehicle (DC), 2.5 uM rosiglitazone (DC RSG), 100 nM AM580 (DC AM). RSG treatment was also combined with 1 uM AGN193109 (DC RSG AGN) in this case two biological replicates was used (RNA was obtained from 2. and 3. individuals).