Uptake of α-Gal A by human kidney ECs.

(A) Dual immunofluorescence staining showing α-Gal A in the glomerulus (G) in a renal biopsy from a Fabry patient who was infused with α-Gal A 2 h before the biopsy. Localization of α-Gal A is observed in the GECs as indicated by co-localization with CD34 (endothelial cell surface marker). Merged high power view demonstrates that α-Gal A is seen in GECs (white arrowheads) and in podocyte (blue arrowhead) in a human glomerulus from a Fabry disease patient. Labeling for α-Gal A is also seen in the proximal tubule (PT). Scale bar, 25 µm. (B) Peroxidase staining showing localization of α-Gal A (green arrowheads) in larger vessel ECs in a renal biopsy from a Fabry patient who was infused with α-Gal A 2 h before the biopsy. Scale bar, 25 µm. (C) Uptake of Alexa-Fluor 546-labeled α-Gal A (red) in cultured human GECs as a function of time at 37°C. At the indicated times, the cells were fixed and analyzed by confocal microscopy. Scale bar, 5 µm. (D) Co-localization (yellow) of α-Gal A (red) and lysosomes (green). A merged image is shown. Scale bar, 5 µm.