Tricorn superose 6 hr 10 300 column

Plasma was fractioned by gel filtration fast-protein liquid chromatography using an AKTA system with a Tricorn Superose 6 HR10/300 column (GE Healthcare Life Sciences, Baie d'Urfe, QC, Canada). Enzymatic assay kits were used to measure total cholesterol (Thermo Fisher Scientific, Ottawa, ON, Canada). IL-6 and tumor necrosis factor (TNF)-α levels were measured by ELISA (Biolegend, San Diego, CA, USA), following the manufacturer's instructions. ] and cells were cultured for 24 h. At the start of each experiment, the medium was changed to fresh Medium 1 containing 3% newborn calf serum (NCS, Life Technologies Inc., Burlington, ON, Canada), or 3% NCLPDS without or with supplementation of different concentrations of human HDL or LDL, without or with addition of apoptosis inducing agents as indicated. Apoptosis inducing agents and concentrations used were: tunicamycin (10 μg/ml); thapsigargin (5 μM); staurosporine (0.3 μM); oxidized (ox) LDL (100 μg protein/ml). Controls contained an equivalent amount of vehicle (0.1% DMSO for tunicamycin, thapsigargin or staurosporine; or saline for oxLDL).