Fig. 3.

CoCl₂ decreases MiTF binding to its own promoter. A, ChIP analysis show that MiTF-CX was associated with MiTF-CX E-box 9 and E-box 3 in untreated cells but not in cervical stromal cells treated with 200 μm CoCl₂ for 24 h. Equal input of DNA from cells treated with and without CoCl₂ was ensured by PCR amplification of DNA extracted from 1/100th of chromatin used for immunoprecipitation of transcription factor-bound DNA (input). Amplification was absent if MiTF antibodies were substituted with IgG or no antibody. B, Cervical stromal cells were treated with CoCl₂ (250 μm) for either 4 or 12 h followed by ChIP analysis and real-time quantitative PCR. *, P < 0.05 compared with time 0.