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. 2019 Oct 7;13(10):e0007742. doi: 10.1371/journal.pntd.0007742

Fig 4. P. brasiliensis Pb18 interaction with an axenic A. castellanii strain.

Fig 4

A) P. brasiliensis and A. castellanii were co-incubated at an MOI of one for one hour at 28°C, and then stained with Giemsa and observed by light microscopy. B) Enlargement of the area depicted in the square region of panel A. C) TEM of the interaction of A. castellanii and Pb18 cells. Incubation was at an MOI of one for six hours at 28°C and then fixed. The black arrow indicates an internalized P. brasiliensis (Scale bar = 2 μm). D) Confocal microscopy. A. castellanii was dyed with DiD-DS (red), while P. brasiliensis cells were labeled first with CMFDA (green), and after the interaction with Uvitex 2B (blue). The arrows show fungal cells inside an amoeba. (Scale bar = 10 μm). E and F) Survival of P. brasiliensis after interaction with A. castellanii. Incubation was at an MOI of two at 28°C for six (E) or 24 hours (F), using the fungus alone as a control. After the interaction amoeba cells were lysed and fungal cells were plated for CFU counting. The figure depicts the results of three independent experiments. The error bars represent the standard error of the mean.