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. Author manuscript; available in PMC: 2008 May 20.
Published in final edited form as: J Biol Chem. 2004 Nov 12;280(2):1236–1240. doi: 10.1074/jbc.M410674200

Fig. 4. Analysis of the association of SNAP-25 phenylalanine mutants with lipid rafts.

Fig. 4

A, immunofluorescence analysis of the subcellular distribution of wild-type SNAP-25 and the F84C and F84S mutants. All three proteins show a strong enrichment at the plasma membrane. B, distribution of proteins between cytosol (C) and membrane (M) fractions purified from PC12 cells. C, membranes prepared from transfected PC12 cells were solubilized in Triton X-100 and fractionated on a discontinuous sucrose gradient as detailed under “Experimental Procedures.” The distribution of the HA-tagged proteins in the recovered gradient fractions was analyzed by immunoblotting using an antibody specific for the HA tag. D, averaged data from six separate experiments. *, F84C displayed an increased level of raft association when compared with wild-type SNAP-25 (p < 0.00005) and F84S (p < 0.035).