FIGURE 7.
The role of palmitate cycling in constitutive localization and activity-dependent redistribution of Gαs-Rluc8. A, net BRET at the plasma membrane and the endoplasmic reticulum generated by Gαs-Rluc8, Gαs-Rluc8 +myr, or Gαs-Rluc8 +myr-palm. PM, plasma membrane; ER, endoplasmic reticulum; GA, Golgi apparatus; EE, early endosomes; LE, late endosomes; RE, recycling endosomes. Bars, mean ± S.E. of n = 5 independent experiments; **, p < 0.001; *, p < 0.05 (one-way ANOVA versus Gαs-Rluc8, Dunnett's multiple comparisons). B, net BRET between Gαs-Rluc8 expressed with unlabeled Gβ1, Gγ2, β2AR, and acceptors localized to membrane compartments in control cells, in cells treated with HDFP (20 μm) for 4 h, and in HDFP-treated cells stimulated for 10 min with isoproterenol (10 μm). Bars, mean ± S.E. of n = 4 independent experiments; **, p < 0.001; *, p < 0.05 (one-way ANOVA versus DMSO (for HDFP) or versus HDFP (for HDFP + isoproterenol), Tukey's multiple comparisons). C and D, net BRET between Gαs-Rluc8 +myr (C) or Gαs-Rluc8 +myr-palm (D) and acceptors localized to membrane compartments in control cells and in cells stimulated for 10 min with isoproterenol. Bars, mean ± S.E. of n = 8–10 independent experiments; **, p < 0.001; *, p < 0.05 (paired t test).