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. 2015 May 3;2015:948131. doi: 10.1155/2015/948131

Figure 4.

Figure 4

Representative confocal fluorescent images of granuloma macrophages stained for different markers. Fragments of splenic granulomas from mouse 2 after 20 days following BCG infection. Nuclei are stained by DAPI (blue signal). Scale bars: 10 μm (a, c, e) and 20 μm (b, d). (a) Mouse granuloma macrophages stained by the LysoTracker Red DND-99 dye (red signal) and mycobacterial LAM-specific antibodies (green signal) show lack of colocalization of BCG-mycobacteria and host cell lysosomes (lack of yellow signal). A single BCG-mycobacterium (green arrow) and replicating BCG-mycobacteria (white arrows). ((b) and (d)) Immunofluorescent localization of CD1d (green signal) and IL-1α (red signal) and IFNγ (green signal) and CD80 (red signal) in granuloma cells, respectively. Colocalization of the markers on the confocal images of cells (yellow signal). ((c) and (e)) The same granuloma fragments as in ((b) and (d)) restained for acid-fast BCG-mycobacteria by the Ziehl-Neelsen method. The same macrophages with replicating BCG-mycobacteria and producing proinflammatory cytokines and leukocyte surface markers are indicated by white arrows on the fluorescent ((b) and (d)) images and by black arrows on the Ziehl-Neelsen ((c) and (e)) images.