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. 2015 Dec 2;7(1):45–55. doi: 10.1080/21505594.2015.1125068

Figure 3.

Figure 3.

Labeling and tracking of acidic organelles in THP-1 cells infected with C. ulcerans. THP-1 cells were incubated with LysoTracker® Red DND-99 for 120 min before cells were infected with C. glutamicum ATCC 13032 pEPR1p45gfp, C. ulcerans 809 pEPR1p45gfp and C. ulcerans BR-AD22 pEPR1p45gfp at an MOI of 10 for 30 min. Extracellular bacteria were killed by the addition of gentamicin and after 2, 8 and 20 h, cells were fixed. Nuclei were stained with DAPI, the cytoskeleton with Alexa Fluor® 647 Phalloidin and micrographs were taken using the confocal laser-scanning microscope Leica SP5 II and analyzed with the LAS software suite. Non-pathogenic C. glutamicum immediately co-localize with acidic compartments (2 h) whereas pathogenic C. ulcerans only show co-localization after longer incubation time (8 h to 20 h). Representative pictures are shown. Scale bars: 10 µm.