Fig. 2

a–d Luciferase Reporter Gene Assays with Caco-2 cells, applying Protocol #3. Caco-2 cells were transiently co-transfected with a human full-length LXRα (a) or LXRβ (b) expression plasmid, the firefly luciferase reporter plasmid ABCA1_Luc and an EGFP expression plasmid, in a ratio of 1:3:1, using Lipofectamine™ LTX. In the case of the mammalian one-hybrid assays, cells were transiently transfected with an expression plasmid encoding a chimeric protein consisting of the yeast Gal4-DBD fused in frame to the hLXRα-LBD (c) or to the hLXRβ-LBD (d), a reporter plasmid containing specific upstream activating sequences (UAS)_Luc, and the EGFP expression plasmid, in a ratio of 5:3:1. Cells were then treated with GW3965 (LXR agonist, 0.1–10 μM) or the vehicle control (DMSO 0.1%). The luciferase-derived luminescence was normalized to the EGFP-derived fluorescence, and set in relation to the solvent vehicle. Bar graphs represent mean ± SD from three independent experiments each performed in quadruplicate. ns not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 versus solvent vehicle (determined by one-way ANOVA with Dunnett post-test)