Fig. 4

Effect of Andro on NF-κB signaling in human breast cancer cells. MDA-MB-231 and MCF-7 cells were cotransfected with a luciferase expression vector containing COX-2 promoter (− 890/+ 9 or − 400/+ 9) 5-flanking fragment (a) and Renilla luciferase reporter (as internal control) for 24 h and then treated with Andro for 48 h. Luciferase activity was determined using a dual-luciferase reporter assay system (b and c). d and e After 48 h treatment, the nuclear extract proteins were prepared and the binding of transactivators (CREB-2, C-Fos NF-κB p65 and p50) to COX-2 promoter probe was analyzed by a streptavidin-agarose pulldown assay. Data were represented as the mean ± S.D. of at least three independent experiments. (*P < 0.05, **P < 0.01, Andro treatment vs vehicle control groups)