FIGURE 1.

Bromopalmitate treatment decreases the association of AKAP79 with lipid rafts. HEK-293 cells stably expressing AKAP79-YFP were treated overnight with bromopalmitate (100 μm in DMSO or control, DMSO only). A, after bromopalmitate treatment, the Triton-soluble extract (Sol) was separated by centrifugation, and the insoluble extract was loaded on sucrose density gradients to isolate lipid rafts. Equal volumes of the fractions and 10 μg of protein from the soluble and total homogenate were immunoblotted for the proteins indicated. PKA cat, PKA catalytic subunit. B, densitometric analysis of blots (mean ± S.E.) expressed as the ratio of immunoreactivity after bromopalmitate treatment over control, for at least four independent experiments. **, p < 0.01; *, p < 0.05. C, confocal imaging of cells expressing AKAP79-YFP treated with 100 μm bromopalmitate and control (DMSO). Scale bars, 20 μm. D, cells treated with bromopalmitate and control were lysed in 0.5 m NaHCO₃ buffer and loaded in a sucrose density gradient. Equal volumes of the gradient fractions and 10 μg of protein from total homogenate were immunoblotted for the proteins indicated.