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. Author manuscript; available in PMC: 2009 Oct 23.
Published in final edited form as: J Biol Chem. 2006 Apr 27;281(27):18351–18362. doi: 10.1074/jbc.M601277200

FIGURE 7. Effects of glucose and WY14643 on L-PK promoter composition.

FIGURE 7

The ChIP assay was used to examine the promoter composition of the rat L-PK and TAT promoters following glucose and WY14643 treatment. Rat primary hepatocytes were treated as described in Fig. 4 and harvested for the ChIP assay. Fragmented chromatin was immunoprecipitated (IP) with antibodies against acetylated histone H4 (Ac-H4) and HNF-4α. DNA was extracted from the immunoprecipitates; 5 μl of 50 μl of purified DNA was used in a PCR to detect the L-PK promoter (−288 to +12 bp) and TAT promoter (−3730 to −3431 bp). Input, 1% of the samples to be used in IP were taken out prior to IP to represent total DNA quantity. A, representative PCR products form the input and products immunoprecipitated with Ac-H4 or HNF-4α antibodies. B and C, PCR products were quantified; mean ± S.D. for three independent studies. Glucose-treated cells (filled circles, solid line); Glucose + WY14643-treated cells (filled box, dashed line). Means ± S.D. for three independent studies. *, p < 0.01 versus glucose-treated cells, Student’s t test.