Figure 2.

Analysis of raft-associated proteins in PC12 cells. Cells were solubilized in 1% Triton X-100 and fractionated on a discontinuous sucrose gradient, as detailed in Materials and Methods. (A) Equal volumes of the recovered fractions were separated by SDS/PAGE and transferred to nitrocellulose for immunoblotting analysis by using antibodies against the indicated proteins. Molecular weight markers are indicated on the Left of all blots. This analysis was repeated on four separate gradients with similar results. Shown is a representative experiment. (B) Comparison of syntaxin 1A and syntaxin 4 association with isolated raft (R) and solubilized (S) fractions from two separate experiments (A and B).