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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 2.

Figure 2

Transient re-expression of Csk in Csk−/− cells results in attenuation of ANP stimulated cGMP production and reconstitution of LPA inhibition. Csk−/− cells grown to 70%–80% confluence in 6 well dishes was transiently transfected with a vector plasmid (Column 1) or plasmids carrying human Csk (Columns 2 and 3) as described in experimental protocols. 16–18 hours after transfection, cells were serum starved overnight and treated for 30 min at 37°C and 5% CO2 with medium containing LPA and 0.5 mM IBMX. This was followed by treatment with 200 nM ANP at 37°C for 3 min. Medium was aspirated, cells were washed once with PBS, lysed with 0.5% Triton X-100 containing 0.5 mM IBMX and cGMP present in the samples was determined. * indicates that the cGMP level from Csk−/−+Csk+ANP is significant different from that from Csk−/−+Csk+ANP+LPA (t-test: p<0.05). The values shown represent the means ± S.D. of three experiments.