2 2 azobis 4 methoxy 2 4 dimethylvaleronitrile

LDL was separated from each subjects’ plasma by single-spin density gradient ultracentrifugation at 100 000 rpm for 40 min at 4°C. The LDL oxidizability was determined by a lag time assay as described^{(22 (link))}. The prepared LDL samples (final concentration of protein: 70 μg/ml) were oxidised by 200 μM 2,2-azobis-4-methoxy-2,4-dimethylvaleronitrile (Wako Pure Chemicals, Osaka, Japan). The kinetics of LDL oxidation were determined by monitoring the absorbance of conjugated dienes at 234 nm with a spectrophotometer (DU800, Beckman Coulter, Indianapolis, IN, USA) at 4-min intervals at 37°C. The lag time for starting LDL oxidation was defined as the time interval between the initiation and the intercept of the two tangents drawn to the lag and the propagation phase of the absorbance curve at 234 nm.

Butyl methacrylate, 4-cyano-4-(phenocarbonothioylthio)pentanoate, poly(ethylene glycol) methyl ether (M_n = 2,000 Da), dicyclohexylcarbodiimide, 4-dimethylaminopyridine, and dichloromethane were purchased from Sigma-Aldrich. 2-(diethylaminoethyl) methacrylate was purchased from TCI Chemicals. 2,2′-azobis(4-methoxy-2,4-dimethylvaleronitrile) was purchased from Wako Chemicals. Pyridyl disulfide ethyl methacrylate was synthesized as described previously (36, 40 (link)). 2′3′-cGAMP was synthesized as described previously (36 (link)). Alexa Fluor 647 Maleimide was purchased from Thermo Fisher Scientific.