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. 2014 Jul 21;4(4):463–473. doi: 10.1021/sb500252a

Figure 3.

Figure 3

Adhesion of E. coli expressing synthetic adhesins to target mammalian cells. (a) Light and fluorescence microscopy images of HeLa-GFP-tm cells grown in culture and infected with EcM1luxSAgfp or EcM1luxSAtir bacteria (MOI 300:1), as indicated. Expression of GFP-tm fusion on the surface of these cells is detected by the green fluorescence emission. (b) Light and fluorescence microscopy images of HeLa-TirM-tm cells grown in culture and infected with EcM1luxSAgfp or EcM1luxSAtir bacteria (MOI 300:1), as indicated. Expression of TirM-tm fusion on the surface of these cells is detected by the green fluorescence emission of its mWasabi domain. Bacteria were stained with anti-E. coli polyclonal serum and anti-IgG-rabbit-Alexa-594 (red). (c) Confocal microscopy images of HeLa-GFP-tm cells infected with EcM1luxSAgfp (top panels) and HeLa-TirM-tm cells infected with EcM1luxSAtir (bottom panels). Infection and staining was carried out as in panels a and b. In addition, cell nuclei and bacterial chromosomes were stained with DAPI (blue). The selected images contain antigen-negative cells (labeled with asterisks) to show the absence of bound bacteria to them.