Detection of AP-1 binding to IGFBP5 promoter at the -1195 T>C. A, Prediction of AP-1 binding site at IGFBP5 promoter -1195 position by TESS (http://www.cbil.upenn.edu/cgi-bin/tess/tess); B, electrophoretic mobility-shift assay with biotin-labeled -1195 T (lanes 1 to 5) or C probes (lanes 6 to 10) and UMSCC-17B nuclear extract. Lanes 1 and 6, probe and nuclear extracts; lanes 2, 7 and 3, 8, probe and nuclear extracts plus 50× unlabeled -1195 T (lanes 2 and 7), or C (lanes 3 and 8) probes; lanes 4 and 9, probe and nuclear extracts and c-fos antibody; lanes 5 and 10, probe and nuclear extracts and rabbit IgG, a relative densitometric assessment of the gel was shown in the bottom panel, with the density of lane 5 setting as 1. The EMSA experiments were repeated once to ensure the consistency; C, ChIP assay using USMCC-17B cells. The binding of IGFBP5 promoter with AP-1 was verified by PCR.