Ampicillin streptomycin

Jurkat (clone E6-1) and HEK-293T cell lines were purchased from the Rio de Janeiro Cell Bank (BCRJ). Jurkat cells were cultured in Roswell Park Memorial Institute (RPMI) medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% ampicillin/streptomycin (Sigma-Aldrich, St. Louis, MO, USA; cat. P4333), and 100 mM of sodium pyruvate (Gibco, Waltham, MA, USA). HEK-293T cells were cultivated in high-glucose Dulbecco’s modified Eagle’s medium (Corning, NY, USA) supplemented with 10% fetal bovine serum, 1% ampicillin/streptomycin, and 100 mM of sodium pyruvate. Cells were maintained in a humid atmosphere with 5% CO₂ at 37 °C and sub-cultured every 2–3 days.

At the time of the collection of blood samples for the study, the blood samples were preserved in ethylenediamine tetra-acetic acid (EDTA), 1% ampicillin/streptomycin, and aprotinin (Sigma-Aldrich, St. Louis, MO, USA) to prevent contamination. Whole blood was centrifuged at 400× g for 15 min at 4 °C to obtain the plasma. Sequential potassium bromide density centrifugation was used to separate LDL from other lipids, such as chylomicrons, very low-density lipoproteins (VLDL), and intermediate-density lipoprotein (IDL) fractions [19 (link)]. LDL was treated with nitrogen and EDTA (5 mM) to prevent ex vivo lipid oxidation.