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Fig. 6
The EDT1-dependent morphogenetic switching pathway plays roles in NADPH oxidase-independent phagocyte migration and virulence.
(A–E) prim25 stage Tg(mpx:GFP)i114 larvae were injected with CAF2-dTomato, edt1Δ/Δ-dTomato, or edt1Δ/EDT1-dTomato C. albicans. Fish were incubated with DMSO or DPI from two hours pre-infection until 4 hpi, and imaged by confocal microscopy. At 4 hpi phagocyte migration and phagocytosis were quantified and fish were sorted into high- and low-responder categories, and at 24 hpi fish were scored for survival. (A) Images are representative of three independent experiments; n = 6 for each fungal genotype and treatment group. Time-lapse images from edt1Δ/EDT1 infections are indistinguishable from wildtype infections but are not shown due to space considerations. Scale bar = 50 μm. (B) Quantitation of total phagocyte response and number of phagocytes with internalized fungi shows the average and standard error from fish pooled from three independent experiments; n = 6 for each fungal genotype and treatment group. (C) Phagocytosis efficiency was measured in fish pooled from three independent experiments; n = 6 for each fungal genotype and treatment group. (D) Survival percentage of low- and high-responders was measured for each of three independent experiments, and the means are shown; n = 70–74 for each group. (E) Low mortality of low-responders infected with yeast-locked edt1Δ/Δ mutant. Means and standard errors of mortality percentages of low- and high-responders at 24 hpi are shown. (B–E) Data represent three independent experiments. Means and standard errors are shown. Differences between groups were assessed by two-tailed Student′s T-test (B, C, and E) or Fisher′s exact test (D). *p<0.05, **p<0.01, ***p<0.001, n.s. no significant difference.