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Fig. 5 | Molecular Medicine

Fig. 5

From: Nitric Oxide Inhibits Endothelial IL-1β-induced ICAM-1 Gene Expression at the Transcriptional Level Decreasing Sp1 and AP-1 Activity

Fig. 5

Sp1 but not EGR-1 binds to the ZIP site of the ICAM-1 promoter after activation of murine aortic endothelial cells with IL-1β. (A) Cells were activated with IL-1β for 2 h and nuclear extracts were prepared. Gel-shift experiments were performed with a consensus sequence corresponding to the ZIP site of the murine ICAM-1 promoter (Table 1) in the absence (lane 1) or presence of 5 µg antibodies specific for Sp1 (lane 2) or EGR-1 (lane 3). Results show that antibodies specific for Sp1 induce a supershift, while anti-EGR-1 antibodies do not. (B) Recombinant EGR-1 (25 ng) was incubated with a consensus sequence corresponding to the ICAM-1 promoter-ZIP site in the absence (lane 1) or presence of 1 µg (lane 2) or 5 µg (lane 3) antibodies specific for EGR-1. Results show, that recombinant EGR-1 is able to bind to the ZIP site and that the EGR-1-specific antibodies induce a supershift.

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Molecular Medicine

ISSN: 1528-3658