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. 2016 Sep 27;6:104. doi: 10.3389/fcimb.2016.00104

Figure 8.

Figure 8

Colocalization between Sau and Spn. Sau and TIGR4 (A–F) or Sau and D39 (G–L) were inoculated together into an eight-well slide and incubated for 1 h (A–C, G–I) or 2 h (D–F, J–L) at 37°C. Biofilms were fixed with 2% PFA and stained with an anti-Sau antibody followed by an Alexa 555-labeled anti-rabbit secondary antibody (red) and then an anti-Spn antibody labeled with Alexa 488 (green). Bacterial DNA was stained by DAPI (blue). Micrographs were taken by confocal microscopy and analyzed using Imaris software. Panels show representative xy optical sections (~0.4 μm each). Bar = 10 μm at right panels and is valid for its corresponding horizontal panels. Vertical panels show specific channels. Arrows point out areas of colocalization between Sau and Spn. (M) Sau colocalized with Spn after 1 h of co-incubation, or free Sau bacteria, were counted in 30 different micrographs. Means were plotted and error bars represent the standard errors. (*), statistical significance (p < 0.001).