Technologies, USA). For this labeling, cells were incubated for 1 h at RT with the fluorescent FAM-YVAD-FMK FLICA reagent following the manufacturer’s instructions (ImmunoChemistry Technologies, USA). The reagent becomes covalently coupled to the active enzyme and is retained within the cell, while the unbound FAM-YVAD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining green fluorescent signal is a direct measure of active caspase-1 enzyme activity. All the negative controls were performed by replacing the respective primary antibodies with isotype and concentration-matched irrelevant antibodies.
