Figure 7.

R. montanensis co-localizes with the lysosomal marker cathepsin D. THP-1-derived macrophages were infected with R. montanensis (MOI = 10). At 60 min or 24 h post infection, cells were fixed, permeabilized, and double stained for immunofluorescence confocal microscopy analysis with NIH/RML I7198 followed by Alexa Fluor 546 (red) to stain R. montanensis, and the monoclonal antibody for cathepsin D followed by Alexa Fluor 488 (green). (A–D) Representative images of a single slice from the z stacks of THP1-derived macrophages at 60 min post infection (A,B) and 24 h post infection (C,D). Each row shows, from left to right, Rickettsia staining, cathepsin D staining, the merged image, and a RGB plot profile illustrating the fluorescence intensity along the magenta arrow. Scale bar = 10 μm. Supplementary Movies 9, 10 represent 360° rotation movie of the 3D projection of the stack images shown in (B) and (C), respectively.