Fig. 4. Subcellular localisation of EndoCL receptor in endothelial cells. (A-D) Intracellular distribution of EndoCL in hDMVECs was assessed by immunofluorescence using LN-1436 antibody and markers of individual cellular structures and organelles: (A) plasma membrane, (B) Golgi, (C) endoplasmic reticulum and (D) lysosomes (see Materials and Methods). The appropriate FITC- (for the detection of hCL; left, first image) or Texas Red- (for the detection of subcellular structures and organelles; centre, second image) conjugated secondary antibodies were used. DAPI was used to counterstain cell nuclei. Colocalized structures (antigens; right, third image) appear in yellow (as indicated by yellow arrows) as determined by overlay of images. Non-colocalised structures appear in green (green arrows) and red (red arrows). Figures are representative of three independent experiments. (For details of a full immunofluorescence screen of intracellular distribution of EndoCL, see supplementary material, Fig. S7.) (E) Localisation of EndoCL at the cell surface in hDMVECs. Reconstructed immunofluorescent confocal pictures of EndoCL in hDMVECs. The en face picture is the collapsed serial of the XY plane confocal image along the Z direction. At the top is the XZ cross-section picture with the apical side facing up and the basal side facing down. To the right of the en face picture is the YZ cross-section picture with apical side on the right and basal side on the left.