Gene expression regulated by AGN193109 and 4-diehtylaminobenzaldehyde (DEAB) in mIMCD-3 cells

All-trans retinoic acid (tRA) is the bioactive derivative of vitamin A that regulates gene expression by activating RA receptors (RAR). By using a reporter mouse model, we recently showed that endogenous tRA/RAR signaling was present in kidney collecting duct cells, and in mouse inner medullary collecting duct cell line (mIMCD-3). In order to unveil target genes of endogenous tRA/RAR signaling in kidney collecting duct cells, whole genome microarray analysis was performed on mIMCD-3 cells treated with AGN193109, a pan-RAR antagonist, and 4-diethylaminobenzaldehyde (DEAB), an inhibitor of tRA synthesizing enzyme. Specificity of gene expression regulation was confirmed by determining the reversibility of the regulation by simultaneous addition of exogenous tRA. In the first set of experiment, mIMCD-3 cells were treated with either vehicle (0.1% ethanol and 0.1% DMSO) or 1 μM AGN193109 or 1 μM AGN193109+0.2 μM tRA. Three independent biological experiments were performed. In the second set of experiment, mIMCD-3 cells were treated with either vehicle (0.1% ethanol and 0.1% DMSO) or 25 μM DEAB or 25 μM DEAB+0.01 μM tRA. Three independent biological experiments were performed. Treatment period for both sets of experiments were 24 hours.