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. 2012 Sep 22;11(12):1898–1912. doi: 10.1074/mcp.M112.019661

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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Fig. 2. — Isolation of detergent-resistant membrane (DRM) and detergent-soluble membrane (DSM) fractions from NB4 cells and the identification of proteins by proteomics. A, successive detergent extraction method to isolate Triton X-100-resistant membranes (DRM) that are rich in lipid rafts. B, quality control Western blots of specific markers for lipid rafts, non-raft membranes, and nuclear proteins. Lyn, a raft-associated kinase, appears only in the DRM fraction; ERGIC-53, an endoplasmic reticulum-Golgi network protein, appears only in the non-raft membranes; and histone-H4 is present only in the nuclear fraction. C, number of proteins identified exclusively in the total cell extract, the DRM fraction, or both fractions. D, cellular compartment ontology identified proteins exclusively in the total cell extract, exclusively in the DRM fraction, or in both fractions. Annotation was obtained with Ingenuity Pathway Analysis software.