Figure 4 | Cell Death & Disease

Figure 4

From: Mycobacterium tuberculosis ESAT-6 is a leukocidin causing Ca2+ influx, necrosis and neutrophil extracellular trap formation

Figure 4

ESAT-6 dependent lysis releases NETs. (a) Neutrophils were treated with 40 μg/ml ESAT-6 and the morphology and propidium iodide (PI) fluorescence (b) assessed by confocal microscopy (white bar=10 μm). Propidium iodide positive cells have an expanded morphology consistent with lysis and extrusion of NET-like structures containing DNA. (c) MPO immunoreactivity of ESAT-6 lysed neutrophils confirms identity as NETs. (d) The mean diameter of necrotic ESAT-6 stimulated neutrophils is significantly greater than necrotic cells from cultures treated with vehicle alone (P<0.001 by T test, cells counted from greater than 10 fields of view). Flow cytometric quantitation of MPO externalisation in ESAT-6 treated neutrophils (e) shows similar high levels of MPO release into the NET as that observed with ionomycin (5 μM) generated NETs (f); anti-MPO FITC (blue) compared with isotype control (green). Representative data from two donors

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