Fig. 7.

DHX9 interacts with p65 and RNA Pol II to active NF-κB-mediated transcription in colorectal cancer cells. A-B Lysates of DHX9-overexpressed or -depleted HCT116 cells were subjected to immunoprecipitation with the anti-DHX9 or anti-IgG antibody, and then incubated with the indicated antibodies using Western blotting. C–D Lysates of DHX9-overexpressed or -depleted HCT116 cells were subjected to immunoprecipitation with the anti-p65 or anti-IgG antibody and subjected to Western blotting. E–G Stable HCT116 cells expressing WT DHX9 and K417R DHX9 were harvested and lysates were used to immunoprecipitation with the anti-DHX9 or anti-p65 or anti-IgG antibody, and then subjected to Western blotting. K417R indicates a negative mutant of DHX9, in which Lys 417 is substituted to Arg that abolishes the ATP-dependent helicase activity. H Stable HCT116 cells expressing WT DHX9 or K417R DHX9 were co-transfected with NF-κB-TATA-Luc reporter plasmid and Renilla luciferase reporter. The luciferase activity of cells was measured after 48-h transfection. The values of firefly luciferase activity were normalized to Renilla luciferase activity. Fold activation were represented as mean ± SEM of three independent transfections. ***P < 0.001, one-way ANOVA with post hoc intergroup comparison by Tukey's test