C. albicans pretreated with SNP+SHAM showed increased uptake by murine macrophages and by zebrafish macrophages in vivo. (A) Wild-type cells were treated with 1 mM SNP–0.5 mM SHAM for 18 h and then washed in phosphate-buffered saline (PBS) and coincubated with J774.1 murine macrophages (3:1 C. albicans/macrophage ratio). Representative examples of uptake after 1 h coincubation are shown. (B) Uptake of cells treated with SNP, SHAM, or SNP+SHAM was quantified by counting at least 200 C. albicans cells per experiment. *, P < 0.001. Five independent experiments were carried out. The percentage of uptake was determined as the number of internalized C. albicans cells relative to the total number of C. albicans cells. (C) Wild-type GFP-labeled C. albicans cells were treated with 1 mM SNP–0.5 mM SHAM for 18 h, washed, and injected into zebrafish larvae. A representative microscopy image of C. albicans (green) being taken up by macrophages (red) in vivo is presented (1, phagocytosed C. albicans; 2, single C. albicans cell; 3, single macrophage). (D) Proportions of phagocytosed C. albicans cells were calculated from images taken from three independent experiments (n = 3). *, P < 0.01. Graphs show means ± standard deviations. Student's t test was used to compare groups.
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