FIG. 6.

Requirement of the −150 SRE for nutritional regulation of FAS in vivo. (A) Diagram showing the mutation introduced at the −150 position within the context of the −444 bp FAS promoter used to generate the −444(−150m) FAS-CAT transgenic mice. (B) RPA was performed on mRNA extracted from the livers of fasted and refed −444(−150m) FAS-CAT transgenic animals to determine the mRNA levels for endogenous FAS and the reporter CAT genes. β-actin mRNA levels are shown as controls. A, B, and C designate the three transgenic lines used in the experiment. (C) ChIP was performed on cross-linked chromatin of livers from fasted and refed −444(−150m) FAS-CAT transgenic mice immunoprecipitated with antibodies against USF and SREBP. DNA was analyzed by PCR with the appropriate primers. LDL receptor (LDLR) promoter is shown as a control. (D) Same as panel B, but the mRNA was prepared from the livers of −444(−150m) FAS-CAT/SREBP double-transgenic mice. Two transgenic lines were used in this experiment. (E) Same as panel C, but the cross-linked chromatin of livers from −444(−150m) FAS-CAT/SREBP double-transgenic mice were used. The results shown are representative of a minimum of three independent experiments for at least two of the founder lines.