Figure 1
From: Link between the causative genes of holoprosencephaly: Zic2 directly regulates Tgif1 expression
Zic2 binds to the Tgif1 enhancer region. (A) Ethidium bromide-stained agarose gel showing the ChIP-PCR analysis. ChIP was performed with anti-Zic2 antiserum or preimmune rabbit serum. The same amount of cell extract was prepared without crosslinking and then purified, and 1% of total DNA mixture was used as a control. (B) DNA footprinting analysis, using DNase I, showing Zic2-binding sites as protected regions indicated at the right side. G indicates the sequence marker, which was digested at the dGTP end. Positions from the transcriptional start site are indicated on the left side. The reaction was performed in the presence or absence of Zic2 protein. (C) The sequence from –1866 to −1649 from the Tgif1 transcriptional start site (+1). (D) EMS assay. To confirm the Zic2 binding sites, an IRD700 end labelled probe (−1866 to −1649) was incubated with Zic2 in the presence or absence of an unlabelled competitor (cp1–cp6). (E) EMS assay showing Zic2 binds to a restricted short fragment probe (−1728 to −1709; Zic2 binding site core).
