Flow cytometry
500,000 to 1x106 PBMC or macrophages were labelled
after Fc receptor blockade with 10% human AB serum by 15 minutes. Then,
cell surface staining was performed with the following monoclonal
anti-human antibodies labelled with different fluorochromes: CD14-FITC
(BioLegend), CD80-PE (BioLegend), CD163-PE (BD Pharmigen), CD206-APC
(Invitrogen), CD36-PerCp (Novus) and ABCA1-PE (Novus). After incubation
and washing, cells were fixed and permeabilized with Foxp3 Fix/Perm kit
(eBioscience) and stained with antibodies against CD68-APC (Invitrogen),
PLIN2-APC (Novus) or IL-10-PE (BioLegend). Cells were analysed with the
FACSCanto II flow cytometer
(BD
Biosciences, Becton Dickinson) and the FlowJo software (Tree Star,
Ashland, OR).