Figure 4.

Inflammasome activation by Fonsecaea pedrosoi hyphae is dependent on caspase-1 and caspase-8. Bone marrow-derived macrophages (BMDMs) were incubated with medium, F. pedrosoi conidia or hyphae for 24 h, and thereafter stained with anti-CD11b-APC antibody and FAM-YVAD-FMK-FITC probe to perform flow cytometry analysis. Caspase-1 activity (indicated by intensity of FAM-YVAD-FMK-FITC binding) of CD11b⁺ cells is represented in histogram and by cell mean fluorescence intensity (MFI) (A). BMDMs obtained from wild-type (WT) or Caspase-1/11^−/− mice were infected with F. pedrosoi hyphae for 24 h (B,C). WT BMDMs were pretreated for 2 h with 50 µM of AC-YVAD-CHO (caspase-1 inhibitor) or Z-IETD-FMK (caspase-8 inhibitor) and then infected with F. pedrosoi hyphae for 24 h (D,E). The supernatant of cell culture assays (B–E) was evaluated for IL-1β and tumor necrosis factor-α by ELISA. Data shown are mean ± SEM and are representative of two to three independent experiments. **p < 0.01; ***p < 0.001, compared to WT-infected untreated cells or between groups indicated by brackets.