Fig 2: Fluorescence imaging of mTagBFP2 fusion constructs.(A–M) C-terminal fusion constructs. For each fusion protein, the linker amino acid length is indicated after the name of the targeted organelle or fusion protein. (A) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs. (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.

Fig 3: FIGURE 2: Time lapse microscopic image series displaying inclusion and plasma membrane rupture in HeLa cells infected with CpoS-deficient C. trachomatis. HeLa cells (ATCC CCL-2) were infected with CpoS-deficient C. trachomatis L2/434/Bu (CTL2-cpoS::bla, 10 inclusion-forming units/cell) and imaged for over 40 hours in 10-min intervals at an Axio Observer.Z1 microscope (Zeiss). The displayed image series (A and B) document the process of necrotic cell death in two selected cells. The morphologic changes in these dying cells suggest that the rupture of the inclusion membrane occurred simultaneously to, or immediately before, the rupture of the plasma membrane. Arrows indicate inclusions.

Fig 4: Inverted-light microscope image analysis of the interaction between HeLa cell line and Acanthamoeba trophozoites from the contact lens paraphernalia and environmental isolates. Images: The arrow shown in the images indicates: (a) The confluent monolayer appearance of the HeLa (ATCC CCL-2) cell line incubated for 24 h; (b) Acanthamoeba trophozoites adapting and attaching to the culture flask; (c,d) the coincubation between HeLa cell monolayer and trophozoites during the adhesion assays. Trophozoites were discovered in close proximity to the surface of epithelial cells as well as beneath the cell layer. Magnification in (a–d) was made at 20×.

Fig 5: Antibacterial activity of PPE in HeLa cells. Survival assay of intracellular S. aureus ATCC 6538 and MRSA strain 2 in HeLa human cells after PPE treatment. HeLa cells (105 cells/well) were infected with staphylococci: (A) S. aureus ATCC and (B) MRSA strain 2 at a multiplicity of infection of 100 treated with GEN and then with PPE at different concentrations or not treated (NT) and re-incubated in Dulbecco’s modified Eagle’s minimal essential medium for the indicated times. After saponin lysis, the numbers of intracellular CFU were recorded. Values are means from at least three independent experiments. The results are shown as the relative number of CFU per well ±SD, and statistical significance was examined using Student’s t-test. Asterisks indicate statistical significance (*** p < 0.001; ** p < 0.01; * p < 0.05).