Fig. 6

CGRP receptor and CGRP recycling. Time course of transient, 100 nM CGRP-induced internalization of CGRP receptors triple positive (yellow) for AA58 (green), RAMP1 (red), CLR-FAP (magenta) (a). CGRP was washed out after 5 min of exposure. Note the appearance of internalized CGRP receptors as early as 10 min after treatment onset and large endosome-like vesicles being present at 60 min after CGRP stimulation (arrowheads). Membrane staining returned after 5 h and was back to control levels after 24 h (a). Time course of sustained, 100 nM CGRP-induced internalization of CGRP (8E11, green) together with the CGRP receptor (RAMP1, red) over 24 h (b). Few double positive, yellow vesicles emerged as early as 10 min after treatment onset (arrowheads), increased in number at 60 min (arrowheads) and in size and labeling intensity at 5 and 24 h (b). Scale bars = 12 μm