Fig. 4.

Transcriptional up-regulation of Twist1 gene by the ALK/Akt/NF-κB axis in UCSs. a The Twist1 promoter sequence containing six putative NF-κB-binding sites. b Various promoter constructs were used for evaluating transcriptional regulation of the Twist1 promoter by p65. Relative activity was determined based on arbitrary light units of luciferase activity normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means ± SDs. The experiment was performed in duplicate. c ChIP assay shows that p65 is bound to Twist1 promoter regions. d Analysis of the indicated protein expression levels by western blot assay in H251-ALK#16 cells after transfection of siRNA against NF-kB/p65. KD, knockdown; con, control; N, nuclear