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Conjugates were then fixed and stained. FITC-CTB-X was used to stain GM1. CD1d was labeled with biotinylated anti-CD1d and then streptavidin-PE. Fixed conjugates were imaged by confocal microscopy. GM1 or CD1d was defined as clustered at the IS if the intensity at the intracellular contact was twice that of the unconjugated membrane. Contacts were defined as areas where co-localization of both mar