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. 2017 Apr 20;114(18):4781–4786. doi: 10.1073/pnas.1619249114

Fig. S7.

Fig. S7.

High-resolution microscopy of S. aureus Newman/pCN57 phagocytosis by Raw 267.4 macrophages. S. aureus Newman/pCN57 (expressing GFP) was added to tissue culture wells containing adherent Raw 264.7 macrophages, supplemented with 5 μg ClyS lysibody or 5 μg PlyG lysibody. Following 1 h of incubation at 37 °C, the wells were washed and macrophages were fixed. Cells were further stained with WGA Alexa Fluor 594 conjugate and imaged using deconvolution microscopy. Images are presented as maximum intensity projections. (A) Representative images of cells treated with ClyS lysibody or PlyG lysibody (scale bar, 5 μm). (B) Serial z sections at 0.6-μm intervals of a single macrophage containing fluorescent staphylococci treated with ClyS lysibody (scale bar, 5 μm). (C) The number of staphylococci in each macrophage was quantified by analyzing the image stack as presented above. The aggregate results from over 100 macrophages per condition are presented. (D) The percentage of intracellular and extracellular bacteria was determined using a similar method; partially phagocytosed bacteria were treated as extracellular.