TIMAP-RACK1 interaction is attenuated by the cAMP/PKA pathway. (A) GST, full-length GST-TIMAP (upper part) or GST-RACK1 (lower part) were immobilized on glutathione-Sepharose and incubated with cell lysates of non treated (ctr), forskolin (50 μM for 30 min) (FRSK) or PMA (1 μM for 30 min) treated BPAEC. The eluted proteins were tested by Western blot using anti-RACK1 and anti-TIMAP antibodies. (B) Endogenous TIMAP or RACK1 was immunoprecipitated from BPAEC lysates after the same treatments described for panel A. IP complexes were probed for TIMAP and RACK1. Shown are representative data of means ± SE from at least 3 independent experiments. Protein levels were quantified by densitometric analysis. Eluted proteins were normalized against total protein levels.