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Fig. 7. Inhibitors of MVBs disrupted inclusion maturation. HEp-2 cells were infected with C. trachomatis E; 1 hour post infection, the inhibitors 3-methyladenine, LY-294002 and U18666A were added to the culture medium at the indicated concentrations. (Upper panel) Fluorescence analysis of the effect of inhibitors on inclusion-size and -maturation assessed at 48 hours post infection. Cells were immunolabeled with the chlamydial-specific anti-hsp60 antibody (anti-mouse Alexa Fluor 594) and 0.5-µm-thick confocal slices revealed significantly reduced inclusion size in the presence of inhibitors. Bar, 20 µm. (Center panel) Electron micrographs of the intracellular bacteria revealed primarily electron-dense EBs in the untreated control cells, but also small inclusions containing RBs in infected cells treated with inhibitors. Bar, 0.5 µm. (Lower panel) A reduction in recovery of infectious Chlamydia was observed in cells cultured in the presence of inhibitors as assessed at 48 hours post infection. Data are presented as mean infectious-forming units of triplicate cultures ± s.e.m.
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