Table 2

Methods for monitoring pyroptosis

IndicatorsMethodReference
Changes in cell morphologyCell swelling, membrane blebbing and rupture, bubble-like protrusionsMicroscopy analysis23,27,48
TEM64,65
SEM21
Automated live cell imager66
GSDM-mediated pore formationliposome leakage method67,68
AFM69,70
Monitoring cell deathCell viabilityMTT/MTS assay71,72
DNA fragmentationTUNEL method73,74
Staining statusAnnexin V/PI staining, SYTOX/7-ADD/EtBr/TO-PRO3 stainingMicroscopy analysis, Flow cytometry23,25,53
Molecular biomarkersCleavage of GSDM family (GSDMB/C/D/E)Western blot
Immunohistochemistry
Immunofluorescence
Q-PCR
23,51,53,75,76
GSDM-Flag
Activation of Caspase-1/3/4/5/11
GzmA and GzmB
Released substances: IL-1β, IL-18, HMGB1, ATP, LDHELISA, ELISPOT27,72,77
Other methodsThe dynamic process of pyroptosis in vivoTwo-photon imaging technology78

TEM, transmission electron microscopy; SEM, scanning electron microscope; GSDM, gasdermin; AFM, atomic force microscopy; GzmA, granzyme A; GzmB, granzyme B.