FIGURE 2.

NFκB p50 is required for fascin protein expression and for Stat3 binding to the fascin promoter. A, MDA-MB-231 cells were transfected with either a control siRNA or NFκBp50 siRNAs and cultured for 4 days. Western blot analyses were performed with antibodies against NFκB p50 or tubulin. B, two rounds of RNAi-mediated knockdown of NFκBp50 was performed (days 1 and 4). On day 7, cells were treated with TNF-α for 3 h followed by Western blot analyses. Autoradiographs of Western blotting analyses were quantitated by ImageJ software, and the results of three experiments + S.D. are shown. C, RNAi-mediated knockdown of NFκB p50 was performed as described in A. Cells were treated with IL-6 for 30 min. ChIP analyses were performed with antibodies against Stat3 or NFκB p50. Results represent two independent experiments.