Fig. 1

MDM efferocytosis of apoptotic granulocytes with flow cytometric gating strategy. Efferocytosis was measured in human MDMs isolated from participants with asthma (n = 19) and healthy controls (n = 10). Human MDMs were co-cultured with PKH-26-labelled apoptotic granulocytes for 90 min at 37 °C in a 24-well plate. The MDMs were harvested and efferocytosis measured by flow cytometry. (a-b) MDMs were gated based on size and granularity using FSC-A vs SSC-A to eliminate debris, doublets and unbound granulocytes. (c) Autofluorescent cells were sub-gated on a FITC plot and (d) the PKH-26 positive MDMs gated on a separate PE plot. (e) The percentage efferocytosis is calculated by the proportion of PKH26-positive MDMs minus the negative control with cytochalasin D. Bars show the means. FSC-A: forward scatter area; FSC-H: forward scatter height; SSC-A: side scatter area