Effect of overexpressing a dominant negative LXRalpha mutant in HepG2 cells

Dominant negative mutatons in nuclear receptors can repress target gene expression, thus having stronger effects than simple loss of gene function. We sought to characterise the effects of a dominant negative mutant (p.W443R) in the hepatoma cell line HepG2 using cell lines stably transfected with wild type and mutant LXR isoforms. HepG2 cells stably tranfected with a construct expressing either a doxycycline inducible wildtype (WT) LXRalpha isoform or a doxycyline inducible dominant negative (p.W443R) isoform (DN) were greated with doxycycline (Dox) to induce expression from the plasmid or vehicle (DMSO) for 24 hours to induce expression of the plasmid, before treatment with either an LXR-agonist GW3965 (GW3965) or Vehicle (Vehicle) for 24 hours.