Fig. 6. Dynamics of internalisation and recycling of EndoCL in response to AM and CGRP. Subcellular distribution of EndoCL in hDMVECs before and after exposure to ligand (100 nM; AM or CGRP) for 30 minute. Cells were processed for immunocytochemistry immediately after exposure, or 2 hours after the complete removal of the agonist from the culture medium (see images labelled 120 mins). Colocalisation of EndoCL with the endothelial cell surface (CD31; top row), early sorting endosome (EEA1; middle row) or lysosome (LAMP1; bottom row) markers (see Materials and Methods). Only merged images are presented. Colocalised structures (antigens; yellow arrows) appear in yellow as determined by overlay of the images. Non-colocalised structures appear in green (green arrows) and red (red arrows). The figures are representative of three independent experiments.