Monocytes were incubated with saturating concentrations of fluorochrome-conjugated mAbs at 4°C for 30 min, and then washed twice in PBS containing 2% FBS and fixed in 1% paraformaldehyde. Data were collected using a FACScan II flow cytometer (BD Biosciences), and data analysis was performed using FlowJo software (BD Biosciences). The fluorochrome-conjugated mAbs against human cell surface proteins CD14, TLR2, and TLR4 and their respective isotype controls were from Bio-Source International and BD Pharmingen.
