Figure 2
PMA induces AP-1 DNA binding to the OPN promoter in HeLa cells. A. HeLa cells were treated with PMA (50 ng/ml) for 2 h. Nuclear extracts were prepared and incubated with 32P-labeled probe containing AP-1 binding site of OPN promoter and analyzed by EMSA. B. For supershift assay, nuclear extracts from PMA-treated HeLa cells were incubated with anti-c-Jun or anti-c-Fos antibody and then analyzed by EMSA. C. HeLa cells were pretreated with TSA (0.5 μM) for 1 h and then with PMA (50 ng/ml) for 2 h. Cross-linked chromatin fragments were immunoprecipitated with anti-p-c-Jun antibody and was PCR amplified using specific primers derived from the region of OPN promoter containing AP-1 binding site. For negative controls, normal mouse IgG was used or specific antibody was omitted.