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. 2006 Sep 27;399(Pt 2):285–295. doi: 10.1042/BJ20060659

Figure 3. In vivo binding of HNF-4α to the FAS gene promoter is increased following the refeeding of carbohydrate to fasted rats.

Figure 3

(A) ChIP assay of FAS promoter using nuclei isolated from liver of fasted or carbohydrate-refed rats. Chromatin fragments immunoprecipitated with HNF-4α antibodies were amplified by PCR with primers spanning the distal carbohydrate region of the rat FAS gene promoter (−7314 to −6980). Immunoprecipitation with normal goat IgG (mock) was used as a negative control and 1% purified input DNA (input) was used a positive control for the PCR reaction. The data shown are representative of three to six individual experiments. (B) Chromatin immunoprecipitated by the HNF-4α antibody was amplified using primers spanning the DR-1 sites of the L-PK and PEPCK promoters. (C) The results of three to six individual experiments were quantified by measuring the density of the PCR products separated on an agarose gel. The numbers are derived from the average density of the PCR products from the refed liver samples compared with the average intensity of those from the fasted liver samples, which are set at 1.0. (D) Whole cell protein extracts from fasted and refed rat liver were analysed by immunoblotting for HNF-4α and actin as described in the Experimental section.