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. 2003 Oct;52(10):1424–1431. doi: 10.1136/gut.52.10.1424

Figure 7.

Figure 7

“Non-raft” localisation of microvillar scavenger receptor class B type I (SR-BI) is not cholesterol dependent. Microvillar membranes were resuspended in 25 mM HEPES-HCl, 150 mM NaCl, pH 7.1, in the presence or absence of 2% methyl-β-cyclodextrin (MβCD) for 30 minutes at room temperature before cooling on ice and solubilisation by 1% Triton X-100 for 10 minutes. After centrifugation at 20 000 g, 30 minutes, the supernatant (NR, the “non-raft” fraction) was collected, and the pellet was resuspended in the above buffer and solubilised by 1% Triton X-100 for 10 minutes at 37°C. After centrifugation as above, the resulting supernatant (R, the lipid raft fraction) and pellet (P, insoluble cytoskeletal proteins) were collected. NR, R, and P samples were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis. After electrophoresis, SR-BI, lactase, and galectin-4 (Gal-4) were detected by immunoblotting.