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Fig. 1. Effect of hypoxia on CYP 2C expression and EET formation in human endothelial cells. (A) Human umbilical vein endothelial cells were transfected with the CYP 2C9 promoter construct and then incubated under normoxic or hypoxic (1% O2) conditions for 1 hour. (B) Time-dependent effect of hypoxia on the expression of CYP 2C mRNA in human endothelial cells as assessed by RT-PCR. (C) Western blot and immunocytochemistry showing the effect of hypoxia on CYP 2C protein levels (green). Expression of HIF-1{alpha} (red) is shown as a positive control. Similar data were obtained in two additional experiments. (D) Effect of the epoxygenase inhibitor MS-PPOH (MS, 10 µmol/l), on the hypoxia-induced increase in 11,12-EET and 11,12-DHET formation. The bar graphs show the mean±s.e.m. of three to six independent experiments; *P<0.05, **P<0.01 vs levels in cells treated with solvent under normoxia (N). Bar, 100 µm.





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