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Fig. 6. Colocalization of VAMP4 and desmoyokin in PC12-27 cells transfected with VAMP4-EGFP. (A-D) Cells transfected with VAMP4-EGFP and immunolabelled in red by anti-GFP and anti-desmoyokin antibodies. Scale bar: 5 µm. (A'-D') The enlargements of the boxes in A-D are shown. The cells in A and A' were fixed while at rest and surface immunolabelled with anti-EGFP without permeabilization. Notice the almost complete lack of red immunolabeling at the surface. The cells in B and C, also non-permeabilized, were fixed after stimulation with ionomycin (3 µM, 5 minutes) and then decorated with anti-GFP and anti-desmoyokin, respectively. The yellowish labelling at the surface documents the incorporation of VAMP4 in the plasma membrane (B') and its colocalization with desmoyokin (C'). (D,D') PC12-27 cells were stimulated, permeabilized and immunolabelled for desmoyokin. Notice that the juxtanuclear EGFP labelling (the TGN) is green, whereas many punctae scattered in the cytoplasm in the proximity of the cell surface are yellowish, documenting the colocalization of VAMP4 and desmoyokin. Only a few green and red punctae are visible in the cytoplasm and also at the surface.
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