T0901317

RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Gibco (New York, NY, USA). Hcy, phorbol-12-myristate-13-acetate (PMA) and Oil-red O dye were supplied from Sigma-Aldrich (St. Louis, MO, USA). SBC-115076 and T0901317 was purchased from Selleck Chemicals (Houston, TX, USA). PCSK9 antibody was obtained from Cell Signaling Technology (Danvers, MA, USA). LDLR, ABCG1, CD68, β-actin and horseradish peroxidase (HRP) goat-anti-rabbit antibodies were both obtained from Abcam (Cambridge, MA, USA). ABCA1 antibody was purchased from Novus Biologicals (Littleton, CO, USA). LXRα was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 22- [- (7- nitrobenz- 2- oxa- 1,3- diazol- 4- yl) amino]- 23, 24- bisnor- 5- cholen- 3β- ol (22-NBD cholesterol) was obtained from Invitrogen (Grand Island, NY, USA). PCR primers were supplied from Sangon Biotech (Shanghai). ApoA-I was supplied from Calbiochem (San Diego, CA, USA) and oxidized-LDL and high-density lipoprotein (HDL) were both obtained from Xiesheng Biotechnology (Beijing).

Authenticated human monocytic THP-1 cells were purchased from the Cell Bank, Type Culture Collection, Chinese Academy of Sciences. THP-1 cells were maintained in RPMI-1640 medium (Gibco) supplemented with 10% FBS, 0.05 mM 2-mercaptoethanol, and 1% penicillin-streptomycin at 37°C in a humidified 5% CO₂ atmosphere. THP-1 cells (2×10⁵ per well) were seeded in 12-well plates and differentiated using 50 ng/ml phorbol 12-myristate-13-acetate (PMA; CS0001, Multi Sciences) for 48 hours to obtain macrophage-like cells. Then, the cells were treated with the LXR agonists GW3965 and T0901317 (Selleck Chemicals) at 1 μM for 24 hours. For gene silencing, differentiated THP-1 cells were transfected with small interfering RNA (siRNAs) targeting NR1H3 (LXR-α), HIF1A (HIF-1α), or scramble control (GenePharma) with GP-transfect-Mate (GenePharma). Oligonucleotide sequences for gene knockdown are listed in Supplementary Table 2. All experiments were performed with mycoplasma-free cells.