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.