Fig. 4

Cholesterol Efflux Assay with Caco-2 cells, using Protocol #5. Caco-2 cells cultivated on filter inserts were loaded with cholesterol by incubation with micelles containing [³H]-cholesterol for 24 h. Hereafter, cells were treated for 48 h with GW3965 (LXR agonist, 5 and 10 μM), T0901317 (LXR agonist, 5 and 10 μM) or the vehicle control (DMSO 0.1%). Finally, cells were incubated with cholesterol-free micelles on the apical side, while being incubated with 1% human plasma on the basolateral side. Effluxed (apical and basolateral supernatant) cholesterol was then measured by liquid scintillation counting. The results in dpm were normalized to 1 ml volume, before being normalized for interexperimental variance. Bar graphs represent mean ± SD from three to four independent experiments. ns not significant, *P < 0.05, **P < 0.01 versus solvent vehicle (determined by one-way ANOVA with Dunnett post-test)