Differential Translocation of Host Cellular Materials into the Chlamydia trachomatis Inclusion Lumen during Chemical Fixation
Fig 1
Mitochondria, ER, and inclusion membranes are found within the lumen of C. trachomatis inclusions.
HeLa cells were infected with C. trachomatis LGV L2, transfected with the indicated plasmids, and fixed at 30 hpi for serial spinning disk laser confocal analysis. Note the presence of markers of the ER, mitochondrial matrix and outer membranes, and inclusion membranes within the inclusion lumen (A, cyan arrowheads). Images portray a single z-section from the center of an inclusion, and inclusions are visually identified as large black centered ovals or outlined with a dashed line. Cellular-localized markers appear saturated because material within the inclusion was often significantly dimmer. (B) The frequency of internalized structures within the entire 3D space of each inclusion was assessed. Plasmids are categorized as markers of the ER, mitochondria, inclusion, cytosol, recycling endosomes, or other as indicated. Within the other category, GFP-GalT localizes to the Golgi, CD63-GFP to MVBs, LAMP1-GFP to lysosomes, and KRphi-mRFP to the plasma membrane. A dashed line at 50% distinguishes between high and low frequencies of intraluminal structures within inclusions. 12–20 inclusions were assessed in each experiment, and the mean ± SEM for three independent experiments is shown. Scale bar represents 5 μm.