Activation of NF-κB activity by GMCSF. (A) HUVECs were incubated in the presence of GMCSF as indicated, followed by preparation of nuclear extracts and electrophoretic mobility shift assay with 32P-labeled NF-κB binding oligonucleotides in the absence or presence of an excess of unlabeled competitor oligonucleotides. (B) HUVECs were transfected with an NF-κB—dependent luciferase reporter construct containing 5 tandem repeats of the NF-κB binding site and an NF-κB—independent β-galactosidase vector as normalization control. One day after transfection, GMCSF or TNF-α was added and cell extracts were prepared after the time points indicated, followed by measurement of luciferase and β-galactosidase activity. The stimulation of NF-κB activity as compared with the vector control is given. Data represent the averages of 2 independent experiments.
