Figure 2.

Direct binding of FOXM1 to the ATX promoter. A. Sequences and positions of putative FOXM-binding sites (#1 site and #2 site) in the ATX promoter and two types of ATX promoter (wild type and mutant type) used in dual luciferase reporter assays were shown. B. Stable PANC-1 cells with FOXM1 knockdown were transfected with 0.2 μg of the wild/mutant type of ATX promoter-luciferase construct, and after 24 hours, the promoter activity was examined by dual luciferase reporter assays. C. Normal PANC-1 cells were co-transfected with 0.2 μg of the wild/mutant type of ATX promoter-luciferase construct and 0/0.1/0.25/0.5 μg of FOXM1-expressing plasmids, respectively, and 24 h after transfection, the promoter activity was examined by dual luciferase reporter assays. D. ChIP assays on putative FOXM1-binding sites of the ATX promoter were performed in PANC-1 cells. A specific anti-FOXM1 antibody was used, normal IgG was used as control, and one percent of the total cell lysates was used for PCR before immunoprecipitation (input control). E. ChIP-qPCR analysis of two FOXM1-binding elements on the ATX promoter was conducted using chromatins immunoprecipitated from PANC-1 cells. *P < 0.05, **P < 0.01.