Primary human monocytes were isolated from the peripheral blood buffy coats by positive selection using MACS CD14+ beads as previously described [50 (link)]. Monocytes were maturated into monocyte-derived macrophages (MDMs) in the presence of 5 ng/ml hGM-CSF (Miltenyl Biotec) treated at days 0 and 2 of maturation. MDMs were used at day 7 of maturation for experiments.
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