After a further 20 min, cells were washed and stained with mouse anti-human CD14–FITC (fluorescein isothiocyanate) (clone 61D3, eBioscience) and mouse IgG (Sigma). Cells were fixed with the paraformaldehyde containing Reagent A (Life Technologies), and data were acquired immediately on a FACSCanto (BD Biosciences) machine. A minimum of 10,000 gated monocytes were acquired for each sample. Fluorescence-minus-one controls were used to set gate thresholds.
