FIG 6.

Gank is upregulated in livers of CUGBP1 S302A mice due to derepression of the Gank promoter. (A) Western blot of Gank in livers of S302A mice 38 weeks after DEN injection. The filter was reprobed with antibodies to β-actin. (B, left) Quantification of protein levels of CUGBP1 as a ratio to β-actin. (Right) Levels of Gank mRNA were determined by qRT-PCR and calculated as ratios to β-actin mRNA. *, P < 0.05. (C) ChIP. C/EBPβ, HDAC1, p300, histone H3K9met, and histone H3K9Ac were immunoprecipitated from chromatin solutions, and the IPs were used for isolation of DNA and PCR with primers covering the C/EBP site within the Gank promoter. (D) Schematic showing the proposed mechanism of Gank promoter activation. (E) Western blotting showing that S302A mice have increased Gank expression at ages of 1 to 8 months (m). (F) Quantification of Gank protein levels as ratios to β-actin.