Figure 5.
Figure 5. AKAP11 binds to free RIα and the RIα holoenzyme. (A) Endogenous AKAP11 was immunoprecipitated from HEK 293 cells in the absence and presence of 20 µM forskolin (FSK)/200 µM IBMX. Samples from the lysate and from the elution of the immunoprecipitation (IP) were immunoblotted (IB) with antibodies against RIα, PKAc, or AKAP11. (B) HEK 293 cells were transiently transfected with control FLAG empty vector, FLAG-tagged RIα, or FLAG-tagged RIα(R211K), an RIα mutant that is defective in cAMP binding. Samples from the lysate and from the elution of the IP were immunoblotted with antibodies against AKAP11, PKAc, or FLAG. (A and B) The lanes marked Lysate were loaded with 5% of the total cell lysates. (C) HeLa cells and Prkar1a−/− MEFs were transiently transfected with GFP-tagged RIα (green) and stained for endogenous AKAP11 (red). Bars, 20 µm.

AKAP11 binds to free RIα and the RIα holoenzyme. (A) Endogenous AKAP11 was immunoprecipitated from HEK 293 cells in the absence and presence of 20 µM forskolin (FSK)/200 µM IBMX. Samples from the lysate and from the elution of the immunoprecipitation (IP) were immunoblotted (IB) with antibodies against RIα, PKAc, or AKAP11. (B) HEK 293 cells were transiently transfected with control FLAG empty vector, FLAG-tagged RIα, or FLAG-tagged RIα(R211K), an RIα mutant that is defective in cAMP binding. Samples from the lysate and from the elution of the IP were immunoblotted with antibodies against AKAP11, PKAc, or FLAG. (A and B) The lanes marked Lysate were loaded with 5% of the total cell lysates. (C) HeLa cells and Prkar1a−/− MEFs were transiently transfected with GFP-tagged RIα (green) and stained for endogenous AKAP11 (red). Bars, 20 µm.

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