Figure 6

Sp1-, C/EBP-β- and NF-κB-DNA binding activities induced by SDF-1 stimulation in DLD-1 cells. (A) Sp1 activation, (B) C/EBP-β activation and (C) NF-κB p65 activation were determined by a TF ELISA assay. (D) The gene silencing efficiency of 48-h transfection of siRNA on Sp1, C/EBP-β, and p65 protein levels of DLD-1 cells. After 48 h of transfection, protein was isolated and the Sp1, C/EBP-β, and p65 expression was analyzed by Western blotting. (E) ICAM-1 mRNA and p537-Luc activity were determined in DLD-1 cells transfected with control siRNA (si-CL), si-Sp1, si-C/EBP-β, or si-p65, and then stimulated with 10 ng/mL SDF-1 for 4 h. All bar graphs represent folds of CL DLD-1 cells, mean ± SEM. *P < 0.05 versus CL. ^#P < 0.05 versus si-CL with SDF-1 stimulation. (F) ChIP assays were performed for Sp1, C/EBP-β, and NF-κB using p65 antibodies.