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. Author manuscript; available in PMC: 2020 May 30.
Published in final edited form as: Annu Rev Immunol. 2020 Jan 31;38:455–485. doi: 10.1146/annurev-immunol-111319-023800

Table 3.

Assays that distinguish membrane-disrupting cell death pathways

Assay Apoptosis Pyroptosis Necroptosis
Count viable surviving colonies
Measure intracellular ATP ↓ (late) ↓ (early) ↓ (early)
Observe cell morphology Cell shrinkage, membrane blebbing Membrane bubbles, cell flattening Cell swelling, cell membrane rupture
Detect the activation of mediators Effector caspase cleavage Inflammatory caspase/GSDM cleavage MLKL phosphorylation
Uptake of cell-impermeable small molecule dyes (SYTOX Green or propidium iodide) Late Early Early
LDH release Late Yes Yes
HMGB1 release No Yes Yes
Annexin V staining Positive Positive Positive
DNA ladder assay, TUNEL assay ++ +/− +/−

Abbreviations: GSDM, gasdermin; MLKL, mixed lineage kinase domain-like pseudokinase; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling.