Automated hs-CRP measurements were performed with immunonephelometry (Beckman Coulter, Milano, Italy), following the manufacture's instruction, and using reagents and calibrators specifically designed for high sensitivity measurement.
PCR and extension reactions were performed in a 96-well plate according to the manufacturer's instructions, using Sequenom reagents.
CD3+, CD4+ and CD8+ were measured by flow cytometry (FACSCount, BD) according to the manufacturer's instructions using standard reagents (BD).
The droplet generation and droplet reading for ddPCR were performed according to the manufacturer's instructions using Bio-Rad reagents.
The GenoType CM/AS (Hain Lifescience GmbH, Nehren, Germany) assays were performed on heat thermo-lysates according to the manufacturer's instructions, using the reagents provided with the kits.
The in situ PLA experiments were performed using reagents and instructions found in commercially available kits from Sigma-Aldrich; Duolink® In Situ PLA® Probe Anti-Rabbit MINUS/PLUS (DUO92005/DUO92002), Duolink® In Situ PLA® Probe Anti-Mouse MINUS/PLUS (DUO92004/DUO92001) and Duolink® In Situ Detection Reagents Orange (DUO92007).
The methylation profile was analysed according to the manufacturer's instructions using Illumina-supplied reagents and conditions.
Droplet generation and droplet reading for ddPCR were carried out according to the manufacturer's instructions using Bio-Rad reagents.
Genotyping was performed by pyrosequencing according to manufacturer's instructions using the PSQ 96 SNP reagent kit (Qiagen, Hilden, Germany) on a PSQ HS96A instrument (Qiagen, Hilden, Germany).
Serum-TRUST and serum-TPPA tests were conducted according to the manufacturer's instructions using the Shanghai Rongsheng Biotech Co TRUST reagent and the Serodia-TPPA reagent from Fujirebio Diagnostics Inc (Malven, Pennsylvania, USA), respectively.
