FIG 5.

NADPH oxidase trafficking coincides with phagosome neutralization and supports depolymerization of DHN-melanin. (A) Representative images of D. discoideum expressing CybA-mCherry after 1 h of infection with FITC-stained conidia of the wild type to assess the phagosomal pH and NADPH oxidase activity simultaneously. The scale bar is 5 μm. (B) FITC-based pH measurements of phagosomes at 1 h p.i. Experiments were performed in 3 biological replicates, and statistically significant differences were calculated with a Bonferroni post hoc test after a two-way ANOVA. (C) FITC-based pH measurements of CybA-mCherry-positive phagosomes of infected cells at 2 h p.i. Experiments were performed in 3 biological replicates. Statistically significant differences were calculated with a t test. (D) H₂O₂-dependent and pH-dependent degradation of synthetic melanin derived from 1,8-DHN or melanin ghosts of wild-type conidia of A. fumigatus. H₂O₂ concentrations are given as wt/vol.