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Fig. 6. Time-lapse photography of auto-aggregates of wild
type (left) or bfpF mutant (right) EPEC bacteria. The wild
type bacteria disaggregate, while the bfpF mutant auto-aggregates
remain fixed. It is believed that BfpF generates the energy required
for pilus retraction, which leads to disaggregation.
Fig. 7. Electron micrograph of purified BfpB showing ring-shaped
mutimers. BfpB is an outer membrane lipoprotein that is believed to
form the channel through which the bundle-forming pilus protrudes.
Fig. 8. The Locus of Enterocyte Effacement (LEE) pathogenicity
island, which consists of 41 open reading frames, is necessary and sufficient
for the attaching and effacing effect. Genes shown in black have
unknown function and are designated either as ORFs or rORFs depending on
orientation. Genes shown in dark blue are esc genes, which
encode homologues of the Yersinia type III secretion system.
Genes shown in light blue are the sep genes, which encode components
of the type III secretion system that lack Yersinia homologues.
The genes in red encodes the outer membrane adhesin intimin. Genes
shown in teal blue encode chaperones of secreted proteins. Genes shown
in green encode esp or other secreted proteins, as indicated. The
positive regulator of the LEE is shown in olive.
Fig. 9. Laser-scanning confocal micrograph of
EPEC interacting with HeLa cells. Bacteria and host cell nuclei are
labeled with DAPI and appear blue. Actin is labeled with FITC-phalloidin
and appears green. Not the large cluster of bacteria toward the left
(localized adherence) and the individual bacteria that are located at the
tip of actin rich pedestals.
Fig. 10. Transmission electron micrograph showing the
attaching and effacing effect. EPEC have effaced microvilli and are
intimately attached to the surface of the HEp-2 cell which responds to
the bacteria by forming the typical cup shaped pedestals that embrace the
Fig. 11. Laser-scanning confocal micrograph of
EPEC interacting with HeLa cells. The image is a composite of a Nomarsky
optical image in which the cells and bacteria are visible, an FITC-phalloidin
stain, in which actin is labeled green, and an affinity-purified EspB antibody
visualized with rhodamine secondary antibody,which labels the EspB protein
red. EspB is visible within the host cell adjacent to bacteria involved
in attaching and effacing.
Fig. 12. A composite image derived from 18 optical sections
taken at 0.5 micron intervals through a HeLa cell that had been infected
with EPEC and stained with affinity-purifed EspB antibodies. The images
were reassembled and are color coded so that those furthest from the camera
are red, those closest are violet and intermediate sections are orange, yellow,
green and blue. These images demonstrate that the EspB protein is distributed
throughout the cytoplasm of the host cell.