Fig. 1.

Rev-erbα Is Expressed in Primary Human Macrophages and Is Regulated by LPS Stimulation, Cholesterol Content, and LXR Activation

A, Rev-erbα mRNA from human aortic SMCs, HUVECs, primary human monocytes and macrophages were quantified by Q-PCR. Nuclear protein extracts were prepared from human primary macrophages and Rev-erbα and β-actin protein levels were measured by western blot analysis. B, Primary human macrophages were stimulated with increasing concentrations of LPS during 24 h. C, Primary human macrophages were cholesterol-loaded with AcLDL (50 μg/ml) for 24 h and subsequently incubated with RPMI 1640 medium with or without apoAI (10 ng/ml). D, Primary human macrophages were incubated with either T0901317 (1 μm) or GW3965 (1 μm). Rev-erbα mRNA levels were quantified by Q-PCR and normalized to cyclophilin mRNA levels. The range of cycle threshold (Ct) measured in cDNA equivalent to 10 ng RNA extracted from primary human macrophages is 24–30 for Rev-erbα compared with 19–21 for cyclophilin. Results are representative of those obtained from three independent macrophage preparations and are expressed relative to the levels in untreated cells set as 1. Each bar is the mean value ± sd of triplicate determinations. Statistically significant differences between treatments and control are indicated (t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001).