Figure 6
Effects of overexpression of AKAP5 on AKAP12-mediated recycling of desensitized, internalized β 2 -adrenergic receptors. HEK293 cells stably expressing β2-AR-GFP or wild-type A431 cells were transiently transfected with an expression vector harboring HA-AKAP5 or the empty vector for 24 h. The cells were untreated or treated with 10 μM isoproterenol for 30 min and then washed free of isoproterenol and allowed to recover for 2 h. Cell surface β2-AR content was detected with the cell impermeant radiolabeled beta-adrenergic antagonist [3H]-CGP12177. A, β2-AR recycling in A431 cells: effects of AKAP5 expression. B, expression of HA-AKAP5 in A431 cells analyzed by immunoblotting and staining with anti-AKAP5 antibody. C, β2-AR recycling in HEK293 cells: effects of AKAP5 expression. D, expression of HA-AKAP5 in HEK293 cells analyzed by immunoblotting and staining with anti-AKAP5 antibody. The results displayed are mean values ± SD derived of triplicates. For panels B and D, the results shown are taken from a single experiment, representative of at least three separate experiments performed on separate cultures of cells. Tabular data from separate experiments was analyzed for variance using the Student's "t" test for significance. # denotes p values > 0.05, i.e. differences that are not statistically significant.