Figure 5. Gankyrin is activated in livers of DEN-treated WT mice via removing of C/EBPβ-HDAC1 complexes from the gankyrin promoter.

A. Expression of FXR, C/EBPβ and gankyrin at early time points after injection of DEN. Western blotting was performed with nuclear extracts isolated at 2, 4 and 7 days after DEN injection. Bottom image. C/EBPβ was IP from nuclear extracts; and HDAC1 was determined in these IPs by Western blotting. IgG; heavy chains of IgGs are shown. B. Expression of C/EBPβ and HDAC1 in FXR/SHP KO mice. Western blotting was performed with nuclear extracts from livers of 4 mice of each genotype. Bottom image: C/EBPβ was immunoprecipitated from liver nuclear extracts of WT and FXR/SHP KO mice and IPs were probed with monoclonal antibodies to HDAC1. C. The gankyrin promoter is activated in livers of DEN-treated WT mice and in livers of FXR/SHP KO mice. ChIP assay was performed with chromatin solutions of WT, DEN-treated WT mice and FXR/SHP KO mice of different ages. D. Activation of FXR in DEN-treated mice prevents elevation of gankyrin. Expression of FXR, C/EBPβ and gankyrin was examined by Western blotting. E. Activation of FXR in DEN treated mice supports C/EBPβ-HDAC1 complexes. C/EBPβ was immunoprecipitated and HDAC1 was examined in these IPs. F. The gankyrin promoter is repressed in DEN-treated mice with activated FXR. ChIP assay was performed as described in legend to figure 5C.