Fig. 6.
Antibody-mediated detection of globo-series neutral GSLs in HBMECs and GMVECs. Lanes a HBMECs, lanes b GMVECs, lanes c human erythrocytes. GSL amounts applied in lanes a and b for anti-Gb3Cer (a) and anti-Gb4Cer (b) overlay assay correspond to 1 × 105 cells, and those in lanes a and b for the orcinol stain (c) correspond to 2 × 105 cells. In lane c amounts of 2 μg (a), 0.2 μg (b), and 20 μg (c) of neutral GSLs from human erythrocytes were used as positive control. Bound anti-GSL antibodies (a, b) were visualized with alkaline phosphatase conjugated secondary antibodies and 5-bromo-4-chloro-3-indolyl phosphate (BCIP) as a substrate. Panel headings Gb3 and Gb4 stand for anti-Gb3Cer and anti-Gb4Cer overlay assay, respectively, and orcinol for orcinol sugar staining. MHC monohexosylceramide