Fig. 6

ATL suppresses the translocation of NF-κB p65/p50 and p300 and the binding of these proteins to the COX-2 promoter. a: After 48 h of ATL treatment, we assessed the binding of p300, p65 and p50 to the COX-2 promoter probe via streptavidin-agarose pulldown assays in U87 cells. b: Correspondingly, chromatin was immunoprecipitated with antibodies against p65, p50 and p300 after treatment with ATL for 48 h; then, we determined the levels of the COX-2 promoter region in the precipitated chromatin by RT-PCR. c: We first separated the cytoplasmic and nuclear proteins and then assessed the expression of p65 and p50 by Western blot analysis with β-actin and Lamin B1 as controls for the sample loading. d: At 48 h after treatment, we also observed the subcellular localization of p65, p50, and p300 and the colocalization of p50 with p65 or p300 by a confocal microscopy analysis