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Endosomal signaling of the receptor for calcitonin gene-related peptide mediates pain transmission

Supporting Information

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  • Download Movie_S01 (AVI) - Live cell imaging of CLR and Cy5-Chol. HEK293 cells were transfected with CLR/RAMP-1. HA-CLR was detected by labeling live cells with HA-Alexa488 antibody conjugate (green channel). Cells were incubated with Cy5-Chol (200 nM). Cells were stimulated with CGRP (50 nM, 30 min) to induce CLR endocytosis. The video was captured on a Leica TCS SP8 Confocal Laser Scanning Microscope and a HCX PL APO 63× (N.A. 1.40) oil objective with a 0.5-AU pinhole (1,800 Hz bidirectional scanning). Stacks of 3 × z sections (0.3 μm per section) were captured every 500 ms. Videos were processed with Huygen’s Deconvolution to reduce noise. (Scale bar, 2 μm.)

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