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Figure 2: Assessment of S. aureus RN6390 internalization using
flow cytometry. Dotted lines indicate the uninfected HEp-2 cell control, and
solid lines indicate HEp-2 cells infected with CFSE-labeled S. aureus ((a)
and (c)) or infected with CFSE-labeled S.
aureus followed by labeling Cy5-conjugated mAb specific for S. aureus ((b)
and (d)). In panels (a) and (b), HEp-2 cells were infected with CFSE-labeled S. aureus without treatment with
lysostaphin. CFSE signal represents HEp-2 cells infected with extracellular
and/or intracellular S. aureus (a).
Cy5 signal represents HEp-2 cells infected with extracellular S. aureus only (b). In panels (c) and
(d), HEp-2 cells were infected with CFSE-labeled S. aureus followed by the treatment with lysostaphin which degrades
staphylococcal cell wall causing a loss of CFSE signal by extracellular S. aureus. CFSE signal represents HEp-2
cells infected with intracellular S.
aureus only (c). This was confirmed by showing the loss of Cy5 signal in
panel (d). Data shown are from a representative experiment which was conducted
three times. |
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