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. Author manuscript; available in PMC: 2014 Jul 21.
Published in final edited form as: Virology. 2010 Sep 28;408(1):89–102. doi: 10.1016/j.virol.2010.08.031

Figure 6.

Figure 6

Scrambling the transmembrane domain prevents VpuEGFP raft association. 293 cells were transfected with vectors expressing either VpuEGFP or VpuTMEGFP proteins. At 48 hours, cells were lysed in ice cold DRM buffer containing 1% Triton X-100, and raft proteins separated from non-raft proteins on discontinuous sucrose gradients by ultracentrifugation as described in the Materials and Methods section. Fractions were collected from the top of the gradient, and fractions from the top (1-3; I) middle (6-7; M) and bottom (10-12;S) were analyzed for the presence of Vpu, Flotillin-1 and transferrin receptor proteins by Western blot analysis using a antibodies described in the Materials and Methods.