Figure 7
Recruitment of MTF-1 to the MREs of the MT-3 promoter after treatment with MS-275. UROtsa parent and transformed cells were seeded at a 1:10 ratio and were grown in the presence of 10 μM MS-275 until they reached confluency. They were then exposed to 100 μM Zn+2 for 4 h and the chromatin was immunoprecipitated with the MTF-1 antibody. PCR was performed with primers specific to each region as specified in Table 2 and Figure 4 to assess the level of precipitated sequences. A. ChIP-q PCR analysis of MTF-1 recruitment to MRE a and MREb. B. ChIP-q PCR analysis of MTF-1 recruitment to MRE-c. C. ChIP-q PCR analysis of MTF-1 recruitment to MREe, MREf and MREg. The amplification value of the immunoprecipitated DNA was normalized to percentage of input (non-precipitated DNA). The determinations were performed in triplicates and the results shown are the mean ± SE.*Statistically significant compared to untreated control cells. Graphs represent real time PCR data whereas ethidium bromide stained gels represent semiquantitative PCR data.