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. 2010 Mar 7;87(3):440–448. doi: 10.1093/cvr/cvq077

Figure 5.

Figure 5

PMCA negatively regulates NO production in endothelial cells. Human PMCA2b or PMCA4b were ectopically expressed in HUVEC by transfection of five μg of the corresponding expression vector (A). NO synthesis was induced by addition of the A23187 ionophore (0.5 μM) 3 min before lyses. NO-dependent cGMP production was calculated in relation to HUVEC transfected with 5 μg of pcDNA3-Empty vector. Mean ± SEM values of six independent experiments are shown. *P < 0.05. (B) Intracellular nitric oxide level in transfected HUVEC was assessed by staining cells with the NO-sensitive dye DAF-FM. NO synthesis was stimulated by addition of acetylcholine (100 μM) for 5 min. Representative microscopy fields of DAF-FM staining in cells are shown in left panels. Mean ± SEM values of fluorescence units measured from 25 different microscopy fields taken from six independent preparations are shown. **P < 0.01.