(A) Cell damage monitored by 51Cr release from RAW264.7 macrophages infected with A. fumigatus wild-type or pksP conidia. Data represent mean ± SD; p < 0.05.
(B) Acidified phagolysosomes (PLs) were stained with LysoTracker red. RAW264.7 macrophage cells were infected with wild-type and pksP conidia for 2 h at MOI = 2. Data represent mean ± SD; p < 0.001.
(C) RAW264.7 cells had simultaneously phagocytosed melanized wild-type conidia (labeled with fluorescein isothiocyanate [FITC], green) and non-pigmented pksP conidia (labeled with Calcofluor white [CFW], blue). Acidified phagolysosomes were stained with LysoTracker red. Scale bar, 10 μm. Lower panel shows a single cell. Scale bar, 5 μm.
(D) A RAW264.7 macrophage had simultaneously phagocytosed a wild-type conidium (FITC labeled) and a pksP conidium (CFW labeled). Localization of cytoplasmic vATPase subunit V1 to the phagolysosomal membrane was monitored by immunofluorescence. Scale bar, 5 μm. See also Figure S1.