Fig. 2.

SiaNAz-labeled molecules are excluded from cholesterol-laden compartments and exhibit partial colocalization with endosomal markers in NPC1-null fibroblasts. (A) NPC1-null fibroblasts were labeled with Ac₄ManNAz and incubated with DIBO and streptavidin Alexa Fluor 568, followed by filipin staining. A representative image is shown. (B) Ac₄ManNAz-labeled control and NPC1-null fibroblasts were incubated with DIBO; costained with monoclonal antibodies to EEA1, CD63, and LAMP1; and then incubated with appropriate secondary antibodies and visualized by confocal microscopy. n = 20–30 cells scored per condition; three independent experiments.