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Figure 1. Expression and localization of GFP-occludin fusion protein in GP8/3.9 rat brain endothelial cells. A–E) GFP-occludin (in green) preferentially localized at the plasma membrane and was recognized by anti-occludin monoclonal antibody (mAb) (in red). Immunoblotting shows that GFP-occludin migrates as a single molecular mass with the expected size of 
90 kDa. GFP-occludin localized at cell-cell contacts and was absent at cell borders lacking apposing cells. F–H) Immunostaining with anti-ZO-1 polyclonal antibody (pAb) (in red) shows that GFP-occludin exactly overlapped with endogenous ZO-1 at cell-cell junctions. I) Expression of ICAM-1, VCAM-1, MHC class II, and vWF as measured by flow cytometry (filled graph: negative control, dashed line: GP8/3.9, solid line: GP8-GFP-occludin) and J) firm adhesion of monocytes were unaffected by GFP-occludin (n=4). Bars: 25 µm
