Fig 4. Binding of Etx correlates with the presence of sulfatide and mono-P IPs in DRM.

(A) After DRM extraction with ultracentrifugation, the fractions from the gradient were pooled as indicated (1/2, 3/4, etc.), and lipids from the pooled fractions were extracted and resolved with thin layer chromatography, as described in the Materials and Methods section. Standards are shown on the left (Std). SM, sphingomyelin; Mono-P IP, monophosphorylated PI. (B) Lipids, extracted under the Blight and Dyer method, were spotted onto a Hybond-C membrane and overlaid with an anti-sulfatide IgG. The presence of sulfatide in DRM fractions was corroborated and correlates with the binding of Etx to lipids from DRM. Lipids were extracted and processed as in Fig 1D.