Under both isotonic and hyperosmotic conditions, a portion of myc-CLN3 is localized in early endosomes and in cholesterol/sphingolipid-enriched plasma membrane microdomain caveolae.

<p>BHK clone 19 myc-CLN3-expressing cells were grown on poly-D-lysine coated coverslips either under isotonic (300 mOsm) or hyperosmotic conditions. Osmolarity was increased at 100 mOsm intervals to 800 mOsm by the addition of NaCl/urea. After being exposed to 800 mOsm for 24 hours, cells were fixed, permeabilized and immunofluorescently stained for the myc-tag (to detect myc-CLN3; green) and for either the early endosomal marker, EEA1 (A) or the caveolae marker, caveolin 1 (B). In the merged images yellow indicates the colocalization of myc-CLN3 with EEA1 (A) or caveolin (B). The white-framed areas of the merged images are enlarged at the right side of the figure to highlight the early endosomal and caveolar localization of myc-CLN3. Arrows in the enlarged images point to colocalizations. Images were taken with a confocal microscope. Scale bars indicate 10 µm.