Caspase Activity Measurement in Cryptococcus

The caspase activity was measured after cell incubation for 24 h, in the presence and absence of synthetic peptides and ITR, according to the methodology described by Qorri and Harless [13 (link)]. After incubation for 24 h, as described above, the samples were washed three times with sterile 0.15 M NaCl and centrifuged (5000× g 5 min at 4 °C) to remove the YPD medium. The samples were washed and centrifuged as described above, and the cells were incubated with 3 μL of CellEvent^® (ThermoFisher, São Paulo, SP, Brasil) for 30 min in the dark at room temperature (22 °C ± 2). Then, the samples were washed and centrifuged again. Finally, the cryptococcal cells were observed under a fluorescence microscope (Olympus System BX60), with an excitation wavelength of 342 nm and emission wavelength of 441 nm.