FIGURE 7.

NOD1 colocalizes with EEA1-labeled endosomes in cells stimulated with EcN or ECOR 12 OMVs. HT-29 cells were incubated with EcN or ECOR12 OMVs (10 μg) for 1 h and colocalization of EEA1/NOD1 was analyzed using laser scanning confocal spectral microscope. NOD1 was stained using anti-NOD1 polyclonal antibody and Alexa Fluor 633-conjugated goat anti-rabbit IgG (red). Endosomes were detected with mouse polyclonal antibody against EEA1 and Alexa Fluor 488-conjugated goat anti-rabbit IgG (green). Images are representative of three independent biological experiments. Colocalization of the red and green signals was confirmed by histogram analysis of the fluorescence intensities along the yellow lines. Analysis was performed as described for Figure 5. Scale bar: 10 μm.