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Rap and PKA activation in living cells. U2OS cells stably expressing Epac1 or Epac2 were stimulated as indicated. The activation of PKA was monitored by a phosphorylation-induced band shift of VASP. Rap•GTP was precipitated from cell lysates and compared to the total Rap levels. control, mock-stimulated; Fsk/IBMX, 15 μM forskolin and 200 μM IBMX to elevate intracellular cAMP levels.
