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. 2010 May 5;285(27):20926–20939. doi: 10.1074/jbc.M109.061622

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — NPR1/GCA promoter activity is inhibited by GREBP overexpression, and this inhibition is dependent on cGMP-RE. A, HEK293 cells were co-transfected with 10 ng of pCMV-βgal, 200 ng of hGCAp-pGL3b, and a combination of varying amounts of pCDNA1-Neo and/or pCDNA1-Neo-GREBP for a total plasmid amount of 500 ng/test. GREBP overexpression was confirmed by sqRT-PCR with specific primers targeting GREBP expressed by pCDNA1-Neo-GREBP. B, HEK293 cells were co-transfected with 10 ng of pCMVβ, 400 ng of pCDNA1-Neo or pCDNA1-Neo-GREBP, and 200 ng of hGCAp-pGL3b or the deleted cGMP-RE form of the human NPR1/GCA promoter hGCAp(ΔcGMP-RE)-pGL3b. All values are expressed as means ± S.E. (error bars) of relative luciferase activity from four different experiments performed in triplicate (n = 12). *, p < 0.05; **, p < 0.0001 versus Neo alone.