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. 2011 Jun 17;6(6):e20894. doi: 10.1371/journal.pone.0020894

Figure 6. Pax6 represses olig2 transcription by binding to the promoter region of the olig2 gene.

Figure 6

(a) Chromatin immunoprecipitation in P4 mouse forebrain lysate indicates that Pax6 binds to the promoter region of olig2. “Input” refers to 0.1%pre-immunoprecipitation input, “w/o Ab” is control without primary antibody, “w/o lysates” is control without lysate, “IgG” is immunoprecipitation with a control IgG, which serves as negative control. (b) Schematics of reporter gene vector. Normal Olig2 promoter region (Olig2-Luc) and Pax6 or HEB binding core sequences mutated promoter region (Pax6 or HEB Binding mut-Luc) are placed upstream of the firefly luciferase gene. (c) Mutated Pax6 or HEB binding site sequences. (d) Reporter assay reveals that Pax6 acts as a transcriptional repressor to olig2. Overexpression of pax6 inhibits the olig2 promoter reporter gene transcription, while the DNA binding site mutant Pax6(5a) R128C does not. A mutation in the Pax6 binding site abolishes the repression effect. Mutation in the HEB binding site abolishes luciferase activity, with or without Pax6. Each result represents a mean ± S.D. of three independent experiments carried out in triplicate. *, by student's t test p<0.05, **, p<0.01.