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Figure 4 | BMC Microbiology

Figure 4

From: CEACAM1 recognition by bacterial pathogens is species-specific

Figure 4

Internalization of Opa CEA -expressing Neisseria gonorrhoeae is only mediated by human CEACAM1. (A) 293 cells were transfected with constructs encoding the indicated human or murine CEACAM1 isoforms fused to GFP or Cerulean. Cells transfected with a GFP-encoding vector served as control. After 48 h, cells were lysed and the expression was determined by Western blotting with a polyclonal anti-GFP antibody. (B) Cells transfected as in A) were infected with Opa-negative (Ngo Opa-) or OpaCEA-expressing N. gonorrhoeae (Ngo OpaCEA) at an MOI of 30 for 2 h. The number of viable intracellular bacteria was determined by gentamicin protection assay. Bars represent mean values ± SEM of three independent experiments done in triplicate. For statistical analysis, samples were compared against control transfected cells by one-tailed Mann-Whitney U-test; *, p < 0.001. (C) 293 cells were transfected with constructs encoding human CEACAM1 isoform containing a short cytoplasmic domain (hCEA1), the corresponding murine isoform (mCEA1) or an empty control vector. Cells were infected with fluorescein-labelled Opa-negative (Ngo Opa-) or OpaCEA-expressing N. gonorrhoeae (Ngo OpaCEA) at an MOI of 30 for 2 h. The uptake index was determined by flow cytometry as described in Material and Methods. Bars represent mean values ± SEM of three independent experiments.

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