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Figure 3


Fig. 3. Interaction of brush border hydrolases with galectin-3 and their association with detergent-resistant membranes. (A) Binding of galectin-3 to hydrolases was assessed by coimmunoprecipitation from small intestinal lysates, using the corresponding antibodies for LPH, DPPIV, SI and APN. Precipitates were subjected to SDS-PAGE and immunoblot analysis with a galectin-3 antibody. When indicated, the coimmunoprecipitation was performed in the presence of galactose (0.3 M) or lactose (0.1 M). (B) Crude membrane pellets of small intestine were extracted with 1% Triton X-100 and separated on a Nycodenz density gradient. Fractions were collected from the top of the gradient (1-12) and subsequently analysed by immunoblot using antibodies against LPH, DPPIV, SI and APN. The distribution of floating flotillin-1 indicates detergent-resistant membrane (DRM)-containing fractions.





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