Fig. 3
EFR3A is present in the DRM fraction and its localization in the DRM fraction is sensitive to the removal of cholesterol from the HeLa cell plasma membrane. A Western blotting of sucrose step gradient fractions obtained after ultracentrifugation of cold 1% Triton X-100 extract (30 min on ice) from HeLa cells. 25 × 106 cells were used. Then, fractions were collected and proteins were precipitated with 10% TCA and analyzed with appropriate primary and secondary antibodies. B Western blotting of the Triton extract from cells treated with 8 mM β-methyl-cyclodextrin shows that localization of EFR3A in the DRM fraction is sensitive to the removal of cholesterol; for comparison, anti-EGFR, and anti-flotillin antibodies reactions are shown. For total protein and cholesterol profiles see Additional file 1: Fig. S3