Neutrophil Apoptosis and Membrane Integrity

Neutrophil apoptosis and loss of plasma membrane integrity were analyzed by flow cytometry (FACSCalibur, BD Biosciences, Oxford, UK or Attune NxT, Thermo Fisher Scientific, Loughborough, UK) of FITC-annexin V (Roche, Welwyn Garden City, UK) and propidium iodide-stained neutrophils^{18 (link)}. Activated caspase-3 was measured using an AF488-coupled antibody specific for cleaved human caspase-3 according to the manufacturer’s instructions. Data were analyzed using FlowJo (TreeStar; version 6.4.7) or FCS Express 7 (De Novo).

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Karmakar U., Chu J.Y., Sundaram K., Astier A.L., Garside H., Hansen C.G., Dransfield I, & Vermeren S. (2021). Immune complex-induced apoptosis and concurrent immune complex clearance are anti-inflammatory neutrophil functions. Cell Death & Disease, 12(4), 296.

Publication 2021

FACSCalibur Attune NxT FITC-annexin V FCS Express 7

Annexin v Antibody specific Apoptosis Caspase Caspase 3 Fitc Flow cytometry Human Neutrophil Plasma membrane Propidium iodide

Corresponding Organization :

Other organizations : University of Edinburgh, Centre de Physiopathologie de Toulouse-Purpan

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Variable analysis

independent variables

Neutrophil apoptosis
Loss of plasma membrane integrity

dependent variables

FITC-annexin V staining
Propidium iodide staining
Activated caspase-3 levels

control variables

Flow cytometry (FACSCalibur, BD Biosciences, Oxford, UK or Attune NxT, Thermo Fisher Scientific, Loughborough, UK)
FlowJo (TreeStar; version 6.4.7) or FCS Express 7 (De Novo) for data analysis

controls

Positive control: Not specified
Negative control: Not specified

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