Raw264.7 cl 0190

We extracted EC from inferior vena cava from BAP31 knockdown mice and purchased human umbilical vein EC (HUVEC) from American Type Culture Collection (ATCC). The two kinds of cells were cultured in ECM (CM-0122; Procell), which contained Ham’sF-12K, 0.1mg/mL heparin, 0.05mg/mL endothelial cell growth supplement (ECG), 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S). We also divided HUVEC into the control group and the experimental group, in which the experimental group added LPS of 1ug/mL and the control group added corresponding dose of phosphate buffered solution (PBS) to observe the effect of LPS on cells. In addition, we also purchased neutrophils (CP-M150; Procell), Raw264.7 (CL-0190; Procell), THP-1 (CL-0233; Procell) and their corresponding media. PMA was added after the THP-1 cells density increased to 1 × 10⁶ so that the concentration of PMA was 50 ng/mL. We also extracted a small number of neutrophils from the peripheral blood of healthy volunteers for the purpose of the experiment (P9402; Solarbio). For more information on volunteers, please refer to Supplementary Table S1. Since neutrophils are terminal cells and are not suitable for long-term culture, cells other than neutrophils are placed in a cell incubator containing 5%CO2 at 37°C.