LDL was aggregated by incubating native LDL (2 mg protein/mL) with sphingomyelinase (Bacillus cereus, catalogue number S9396-25UN, Sigma) at 10 mU/mL (Walters and Wrenn, 2010 (link)) until the attenuance (absorbance plus light scattering) at 680 nm increased from about 0.0017–0.027. sphingomyelinase-aggregated LDL (SMase-LDL) was dialysed against a phosphate buffer containing EDTA and sterilised by membrane filtration (0.45 μm).
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