Microphthalmia Transcription Factor Regulates Pancreatic β-Cell Function

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FIG. 6.
FIG. 6.

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.

This Article

  1. Diabetes vol. 62 no. 8 2834-2842