Figure 6.
CLN3 localization to TGN and membrane microdomains. A, MBEClacZ/lacZ were transfected with empty control plasmid (pBUD-mcs) or a CLN3 expression plasmid (pBUD-CLN3). Twenty-four hours post-transfection cells were fixed, permeabilized, and dual stained for CLN3 and a marker for GM1 microdomains (A488-CTB), TGN (WGA or TGN46), or cis-Golgi (GM130). Stained cells were analyzed by confocal microscopy. Representative images are shown. Scale bars, 20 μm. B, MBEClacZ/lacZ transfected as in A were extracted with carbonate and fractionated over a discontinuous sucrose gradient. Fractions from top to bottom were analyzed by Western blot for CLN3 and by dot blot for GM1.