Mitf binds to and regulates Pax6 expression in β-TC6 cells. Pancreas-specific Pax6 expression is partially regulated by upstream
regulatory elements (summarized in ref. 25). A: The pPax6P0-LUC construct contains the EE and PE elements. B: ChIP analysis was performed on β-TC6 cells transfected with an expression vector containing FLAG-tagged Mitf. The anti-FLAG
precipitated chromatin was analyzed by nonquantitative PCR using primers specific for Pax4, Pax6EE, Pax6PE, and PEPCK control region sequences. C: ChIP analysis of α-TC6 cells transfected with FLAG-tagged Mitf. PCR analysis with Pax4, Pax6EE, Pax6PE, and Runx2 control region sequences. Control PCR reactions were run with input chromatin (1:100 dilution), and DNA was obtained after
precipitation with mouse IgG or no DNA. ChIP analysis was repeated from at least three independent chromatin preparations.
D: Dual luciferase reporter assays show that pPax6P0-LUC activity is enhanced by cotransfection with Pax6 in HEK293 cells but
repressed by cotransfection with Mitf.