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. 1999 Nov 9;96(23):13357–13362. doi: 10.1073/pnas.96.23.13357

Figure 6.

Figure 6

Effect of dexamethasone on NOS III mRNA stability and NOS III promoter activity in EA.hy 926 cells. In a, EA.hy 926 cells were preincubated without (●) or with 100 nM dexamethasone (□) for 18 h. Then, the inhibitor of transcription actinomycin D (10 μg/ml) was added to the culture medium. RNA was prepared 0, 12, 18, 24, and 48 h thereafter, and NOS III mRNA was determined by RNase protection assays. The NOS III mRNA levels in both groups were set at 100% at the time of addition of actinomycin D (0 h). Values are means ± SEM of three independent experiments. b shows EA.hy 926 cells stably transfected with pNOS III-3500-Hu-Luc-neo (containing a 3.5-kilobase NOS III promoter fragment cloned before a luciferase reporter gene). Stable cells either were kept untreated [control (Co)] or were incubated with dexamethasone (10–1,000 nM) for 24 h. Then, the cells were lysed, and luciferase activity was determined. The luciferase activity was taken as a measure of NOS III promoter activity. Bars represent means ± SEM of three independent experiments. Asterisks indicate significant differences from untreated cells (**, P < 0.01; ***, P < 0.001 by ANOVA and Fisher’s PLSD test).