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. 2006 Mar 24;103(14):5379–5384. doi: 10.1073/pnas.0601026103

Fig. 1.

Fig. 1.

Effect of endogenous NO on HIF-1α accumulation at different O2 concentrations. HIF-1α protein levels were detected by Western blotting of nuclear extracts from HUVECs, HEK293 cells, and smooth muscle cells (A) or human microvascular endothelial cells (B) exposed for 8 h to the indicated O2 concentration in the absence or presence of l-NMMA (1 mM). (C) HIF-1α protein levels from nuclear extract of HUVECs exposed to 1.5% O2 in the presence or absence of l-NMMA and myxothiazol. (D) HIF-1α DNA-binding activity in nuclear extracts from HUVECs exposed to O2 concentration as in A. The values represent the mean ± SEM from three independent experiments. (E) HUVECs infected with short-hairpin RNA eNOS exhibited markedly reduced eNOS expression levels after 2 weeks when compared with wild-type cells. GAPDH was used as protein loading control. (F) HIF-1α protein levels from nuclear extract of silenced eNOS cells exposed to different O2 concentrations. Values in brackets are the estimations from densitometry analysis.