Fig. 2. Distribution of VAMPs and annexin-2 in PC12-27 cells, relative to the enlargeosome marker desmoyokin. The four series of deconvolved confocal images, obtained from PC12-27 cells that were permeabilized before immunolabelling, illustrate: (left) the distribution of VAMP4, VAMP7 and VAMP8 (A-C, respectively), and of annexin-2 (D); (middle) that of desmoyokin; (right) the merge of the two corresponding images. The VAMP4 results, obtained with a monoclonal antibody raised in our laboratory, are representative also of those obtained with another monoclonal antibody and with two polyclonal antibodies. All the VAMP proteins and annexin-2 appear spread in the cytoplasm, with VAMP4 concentrated preferentially in the perinuclear area (TGN) and annexin-2 showing a positivity at the periphery of the cell. The merge images reveal a significant colocalization of desmoyokin in the cytoplasmic punctae that were positive for VAMP4 and annexin-2 (A,D). No significant level of colocalization is visible for VAMP7 and VAMP8 (B,C). Scale bar: 5 µm.