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. Author manuscript; available in PMC: 2008 Aug 24.
Published in final edited form as: Biochemistry. 2006 Mar 14;45(10):3396–3403. doi: 10.1021/bi052513u

Figure 3.

Figure 3

The catalytic activity of Csk is essential for its regulation of receptor guanylyl cyclase activity. A. Csk−/− cells stably expressing the CskR318A mutant were grown to 80% confluence in 6 well dishes. Following serum starvation overnight, cells were treated with medium containing LPA and 0.5 mM IBMX for 30 min at 37°C in the presence (▼) or absence (■) of 50 mM imidazole. This was followed by treatment with indicated concentrations of ANP for 3 min at 37°C. Cells were then washed once with PBS, lysed with 0.5% Triton X-100 containing 0.5 mM IBMX and cGMP content present in the samples was determined. B. Treatment of Csk−/− cells with imidazole (Column 3) or transient expression of CskR318A in Csk−/− cells (without imidazole addition) (Column 4) did not lead to significant changes in ANP-stimulated cGMP production. Data are representative of three similar experiments.