Fig. 2.

Effect of H₂O₂ on transcription factor binding to the AP-1 cis-element in the human eNOS promoter. EMSA analyses were performed using a biotinylated AP-1 oligonucleotide corresponding to −661 of the human eNOS promoter. The biotinylated oligonucleotide was incubated for 20 min with nuclear extracts from FPAECs treated with 100 μM H₂O₂ for 2 h. The reaction was run on a 5% native polyacrylamide gel and transferred to a nylon membrane, and the biotinylated oligonucleotide was detected by using a streptavidin-horseradish peroxidase antibody and chemiluminescent substrate. Arrow indicates the position of the AP-1 complex. Lane marked Free is biotinylated oligonucleotide without any extract. The bar graph shows relative changes in band intensity for FPAECs exposed to H₂O₂ for 2 h. Band intensities were calculated by densitometric analysis. Binding is expressed as fold changes relative to untreated control (Cont). The values expressed are means ± SE, n = 6. *P < 0.05 vs. untreated.