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Figure 7 | Veterinary Research

Figure 7

From: Occludin and collagen IV degradation mediated by the T9SS effector SspA contributes to blood–brain barrier damage in ducks during Riemerella anatipestifer infection

Figure 7

SspA binds to occludin. A SspA expression in 293 T cells (Lane 1: lysate of 293 T cells transfected with the plasmid pcMV-flag; Lane 2: lysate of 293 T cells transfected with the plasmid pcMV-sspA-flag). B SspA has hydrolytic activity towards gelatine (Lane 1: 2 µg of recombinant rSspA protein; Lane 2: lysate of 293 T cells; Lane 3: lysate of 293 T cells transfected with the plasmid pcMV-flag; Lane 4: lysate of 293 T cells transfected with the plasmid pcMV-sspA-flag). C Occludin expression in 293 T cells (Lane 1: lysate of 293 T cells transfected with the plasmid pcDNA-myc; Lane 2: lysate of 293 T cells transfected with the plasmid pcDNA-occludin-myc). D The subcellular distribution of SspA (red) and occludin (green) was examined by immunostaining with anti-FLAG-C or anti-MYC-C antibodies. Nuclei were stained with DAPI (blue). Merged images showing SspA/occludin/DAPI fluorescence. E Immunoprecipitation of SspA and occludin. 293 T cells were co-transfected with pcMV-sspA-flag and pcDNA-occludin-myc. The cell lysates were incubated with anti-Flag magnetic beads and subjected to immunoprecipitation using an anti-MYC-C tag polyclonal antibody. The precipitated proteins and whole-cell lysates were analysed by Western blotting.

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Veterinary Research

ISSN: 1297-9716