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Figure 6 Effects of intravenous Ang-II injection on nuclear factor-kappa B (NF-kB) activity and expression of proinflammatory cytokines in heart tissue. (A) Representative electrophoretic mobility shift assay experiment. From each sample, equal amounts of nuclear protein in heart tissue were obtained and mixed (n = 5 in each group); 8 µg of the mixed proteins was used for the experiments. Lane 1 = wild-type (WT) with phosphate buffered saline (PBS) injection; lane 2 = knock-out (KO) with PBS injection; lane 3 = WT with Ang-II injection; lane 4 = KO with Ang-II injection. (B) Densitometric analysis of the results expressed as arbitrary units of mean values ± SEM (5 animals from each group). Mean NF-kB activity in the WT group with PBS injection is represented as 1. At 30 min after the injection of Ang-II, NF-kB activity was significantly increased in WT but not KO mice. (C, D) Gene expression of proinflammatory cytokines in heart tissue harvested 90 min after the injection of Ang-II or PBS. The cytokine mRNAs were corrected by the amount of GAPDH mRNA. At 90 min after the injection of Ang-II, the expressions of TNF-alpha (C) and IL-1-beta (D) were significantly increased in WT but not KO mice (n = 5 each). *p < 0.01 vs. WT with PBS injection. #p < 0.01 vs. KO with Ang-II injection.
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