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Fig. 2. Continuous expression of the SOD1 G93A mutant selectively affects the surface distribution of GLT-1 in MDCK cells. (A,B) Forty-eight hours after transfection, the cells were fixed and triple-stained with antibodies against GLT-1, EAAC1 or GLAST (red); SOD1 (green); and ß-catenin, a marker of the lateral junctional domain (blue). Confocal horizontal (A) and vertical (B) sections show the distribution of wild type and chimeric transporters in SOD1- and G93A-expressing cell lines. The confocal merges show surface (apical and basolateral) localisation of the GLT-1 and EAAC-GLT in SOD1-expressing cells, and their intracellular accumulation in G93A-expressing cells. The surface distributions of the neuronal EAAC-1 transporter (apical surface) and the glial GLAST (basolateral surface) are retained in the G93A-expressing MDCK cell lines. The arrows in A indicate perinuclear staining, and the arrows in B the apical cell surface. Bar in A, 5 µm; B, 10 µm.

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