Cs0001

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.

RAW 264.7 (a mouse-derived macrophage cell line) and THP-1 (a human monocytic leukemia cell line) cells were purchased from Procell Life Science Technology Co. Ltd. (Wuhan, China). Cells were cultured at 37 °C with 5 % CO_2. The culture medium for RAW 264.7 cells was DMEM supplemented with 10 % FBS and 1 % penicillin-streptomycin. RAW 264.7 cells were seeded in a complete culture medium in 6-well plates at a density of 5 × 10⁵ cells/well. After overnight incubation, cells were treated with LPS (Solarbio, L8880) (1 μg/mL) for 24 h.
The culture medium for THP-1 cells was RPMI 1640 supplemented with 10 % FBS, 1 % penicillin-streptomycin, and 0.05 mM β-mercaptoethanol. THP-1 cells were also seeded in 6-well plates at a density of 5 × 10⁵ cells/well. THP-1 cells were differentiated into macrophages by incubating with 100 ng/mL phorbol 12-myristate 13-acetate (MultiSciences, CS0001) for 48 h. Then, cells were treated with LPS (1 μg/mL) for 24 h.
The other RT-qPCR steps were the same as those used in the clinical cohort. Table S2 lists the primer pairs used in the experiment.