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. 2014 Jan 3;9(1):e84704. doi: 10.1371/journal.pone.0084704

Figure 5. NET formation induced by complement-mediated phagocytosis against the invasion of C. albicans.

Figure 5

(A) Serum-treated or non-treated C. albicans were added to neutrophil-like differentiated HL60 cells (the ratio of C. albicans to cell; 5∶1) in the presence of Sytox Green for 3 h at 37°C. Left panel shows representative fluorescence images and right panel shows frequency of Sytox green-positeve cells. Green shows Sytox Green staining. (B) Serum-treated C. albicans were added to neutrophil-like differentiated control cells and Rab27a-knockdown cells in the presence of both Hoechst 33342 and Sytox Green for 3 h (The ratio of C. albicans to cell; 5∶1). Left panel shows representative fluorescence images. Blue shows Hoechst 33342 staining and Green Sytox Green. Right panel shows the percentage of the cells in the individual stage (black: Hoechst 33342–positive; white: Sytox Green-positive and decondensed; shaded area: Sytox Green-positive and cloud-like shape). (C) Serum-treated C. albicans were added to neutrophil-like differentiated control cells and Rab27a-knockdown cells in the presence or absence of DPI for 3 h. Frequency of Sytox green-positive cells are shown. In (A) and (B), Scale bars indicate 10 µm. In (A), (B) and (C), the results of Rab27aKD clone 1 and Control clone 1 are shown and the data are the mean with SD from three independent experiments. Asterisks (*) mean that the difference is statistically significant (p values <0.01).