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. 2010 Nov 11;286(2):1499–1507. doi: 10.1074/jbc.M110.189464

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Effect of MCD on membrane distribution of Dsc2 and Oly. A, cells were grown in control and MCD media and labeled with Oly. In the control treatment, Dsc2 overlapped with Oly (arrows, upper panel). Oly labeling completely disappeared from the plasma membrane after cholesterol depletion (lower panel), but the Dsc2 distribution did not disappear. Scale bar, 10 μm. B, depletion of membrane cholesterol with MCD released Dsc2 from DRMs and non-DRMs (MCD). Dsc2 distribution in DRMs was returned when MCD-cholesterol was applied to cells (MCD-CHOL). Treatment with cholesterol oxidase (CO) completely released Dsc2 from DRMs. C, effect of MCD on membrane distribution of Dsg2 is shown. Fraction 5 and DRM fractions 6 and 7 were pooled from sucrose gradients after centrifugation of Triton X-100-lysed cells. Dsg2 is released from fraction 5 and from DRM fractions 6 and 7 compared with control fractions.