FIG. 4.

Binding of CFA-1 fimbriae of ETEC to pure glycosphingolipids on thin-layer chromatograms. Chemical detection by anisaldehyde (A) and an autoradiogram obtained by binding of ¹²⁵I-labeled CFA-1 fimbriae (B) are shown. The glycosphingolipids were separated on aluminum-backed silica gel plates with chloroform-methanol-water (60:35:8, by volume) as the solvent system, and the binding assays were performed as described in Materials and Methods. Autoradiography was performed for 12 h. Lanes: 1, neolactotetraosylceramide (Galβ4GlcNAcβ3Galβ4Glcβ1Cer) (2 μg); 2, sialyl-neolactotetraosylceramide (NeuAcα3Galβ4GlcNAcβ3Galβ4Glcβ1Cer) (2 μg); 3, Le^x pentaglycosylceramide [Galβ4(Fucα3)GlcNAcβ3Galβ4Glcβ1Cer] (2 μg); 4, sialyl-Le^x hexaglycosylceramide [NeuAcα3Galβ4(Fucα3)GlcNAcβ3Galβ4Glcβ1Cer] (2 μg); 5, Le^a pentaglycosylceramide [Galβ3(Fucα4)GlcNAcβ3Galβ4Glcβ1Cer] (2 μg); 6, sialyl-Le^a hexaglycosylceramide [NeuAcα3Galβ3(Fucα4) GlcNAcβ3Galβ4Glcβ1Cer] (2 μg).