Fig. 3.

Extracellular ROS accumulate in the presence of sod5Δ/Δ cells. A. Superoxides measurement by lucigenin-dependent chemiluminescence at 37°C over a 90 min period [relative luciferase units (RLU) under the curve]. Stimulation of BMDMs with either the wild-type (CA-IF100) strain, or the sod4Δ/Δ (CA-IF015), sod5Δ/Δ (CA-IF019) mutant strain or the sod5Δ/Δ::SOD5 revertant (CA-IF027) (MOI 5:1). *Infection with sod5Δ/Δ yields 3.2 ± 0.21 times more superoxides than with wild-type C. albicans**P > 0.005. B. Extracellular ROS measurement by isoluminol-dependent chemiluminescence at 37°C in 2.5 min intervals over a 90 min period [relative luciferase units (RLU) min⁻¹ per 1000 cells]. Stimulation of BMDMs with either the wild-type (CA-IF100) strain or the sod5Δ/Δ (CA-IF019) or sod6Δ/Δ (CA-IF023) mutant strains (MOI 5:1). Quantification of the total ROS release between 30 and 70 min (striped area) by calculating the area under the curve. *Infection with sod5Δ/Δ yields 10 ± 0.5 times more extracellular ROS than with wild-type cells. **P > 0.001. C. Intracellular ROS production in response to the phorbol ester PMA, wild-type (CA-IF100) strain or sod5Δ/Δ (CA-IF019) mutant strain was measured by FACS analysis using H₂DCF-DA staining of BMDMs after 15 min (a) or 40 min (b) of infection. A–C. Results of one experiment per condition are shown. All data were reproduced in at least three independent experiments. Statistical significances were calculated using a two-tailed Student's t-test.