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Figure 2 | BMC Biochemistry

Figure 2

From: Protein kinase A type I activates a CRE-element more efficiently than protein kinase A type II regardless of C subunit isoform

Figure 2

Activity of PKA R and C subunits expressed in 293T cells. (A and B) 293T cells were left untransfected, transfected with empty vector (vector) or with increasing amounts (0 - 5600 ng) of either pDeCα1 (Cα1, —) or pDeCβ2 (Cβ2, ------). After 24 hours cells were harvested, homogenized and all cell extracts adjusted to 1 mg total protein/mL. PKA activity was determined as catalytic activity against Kemptide in the presence of 7.14 μM cAMP (A) and Luciferase activity at 560 nm (B). Data points represent enzyme activity and relative luciferase activity, respectively, +/- SD, n = 3. (C and D) 293T cells were left untransfected, transfected with empty vector (vector) or with increasing amounts (10 - 1280 ng) of either pDeRIα or pExRIIα. Levels of R subunit expression were monitored as [3H]-cAMP-binding and R subunit immunoreactivity against RIα (C, clone 4D7, 1: 300 dilution) or anti-RIIα (D, 1 : 400 dilution) after SDS-PAGE separation of 25 μg total protein per lane in 12.5 % gels. R subunit activities are given as cpm +/- SD (n = 3). The apparent molecular weight of protein recognized is indicated by arrows and protein identity RIα (49 kDa) and RIIα (52 kDa), given by arrows to the left. One immunoblot out of three independent experiments is shown.

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