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Fig. 6. Colocalization of the palmitoylation-deficient mutants of claudin-14 with the lysosomal membrane protein LAMP-2. Tet-off MDCK cells transfected with the palmitoylation-deficient mutant of claudin-14 were plated on filters and induced to express the transgene. As demonstrated in Fig. 5, the 4S mutant of claudin-14 was found at cell borders and in large intracellular aggregates (top right). ZO-1 (top left) was concentrated at the apical cell membrane, but was also found in large intracellular aggregates. LAMP-2-positive structures (bottom left) did not appear to be affected by induction of mutant protein (not shown). Immunofluorescent analysis revealed colocalization (bottom right) of the intracellular fraction of claudin-14 4S mutant (red) with LAMP-2 (green). Staining with antibodies against other intracellular compartments (early endosomes, Golgi) were negative for colocalization with mutant claudin-14. Scale bar, 10 µm.





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