1% NDS. Tissue was incubated overnight at 4°C in primary antibody, washed with 0.1 M PBS with 0.3% Triton X-100 plus 1% NDS, and then incubated for 2 hrs in secondary antibody at RT. Tissue was then washed with 0.1 M PB, and coverslipped with Fluoromount-G mounting medium (Southern Biotech). Images were acquired with a Leica TCS SPE confocal microscope. We used the following antibodies and reagents: sheep anti-CGRP (Abcam, 1:1,500); IB4 (Sigma-Aldrich, 1:1,000); guinea pig anti-Substance P (gift from J. Maggio, 1:10,000), Alexa Fluor 488 donkey anti-sheep; Alexa Fluor 555 streptavidin, Alexa Fluor 647 donkey anti-guinea pig.
