Two Distinct Coagulase-Dependent Barriers Protect Staphylococcus aureus from Neutrophils in a Three Dimensional in vitro Infection Model (original) (raw)
Figure 3
Localization of Coa and Emp to S. aureus pseudocapsules.
Coa and Emp were detected in pseudocapsules by immunocytochemistry using rabbit antibodies specific for coagulase (anti-Coa, A) or Emp (anti-Emp, B and C). The pseudocapsule of the coa mutant is more irregularly shaped than its wildtype counterpart, illustrated by anti-Emp staining (C). Staphylococci were grown in 3D-CoG/Fib for 16 h and then fixed with 4% paraformaldehyde. Primary antibodies were detected by Alexa Fluor 555 anti-rabbit antibodies. Staphylococci were detected by staining DNA with DAPI (A1, B1, C1) and N-acetyl-glucosamine with FITC-Lectin (T. vulgaris) (A2, B2, B3). Staining of Coa and Emp was specific as the coa and vwbp emp mutant were not stained with the respective antisera (data not shown). Scale bar 7.5 µm. The images are representative of three independent experiments.