Figure 2. Feeding-induced S262 phosphorylation and K237 acetylation of USF-1.
(A) USF-1 immunoprecipitates using monoclonal anti-USF-1 antibodies was Western blotted with polyclonal anti-USF-1 or anti-P-USF-1 antibodies. Immunoblotting with anti-P-USF-1 in the presence of peptide or with pre-immune serum are shown as controls. (B) ChIP for indicated proteins binding to the −444 FAS-CAT promoter in mouse liver. (C) ChIP using anti-FLAG antibodies (top) for WT USF-1 and S262 USF-1 mutant association to the FAS promoter in 293FT cells. The FAS promoter activity (bottom) in cells transfected with −444 FAS-Luc and WT USF-1 or S262 USF-1 mutants was monitored. Immunoblotting for protein levels of WT, S262 USF-1 mutants (insert) and FAS are shown. (D) Immunoprecipitated USF-1 was Western blotted with indicated antibodies. (E) ChIP for binding of indicated proteins to the −444 FAS-CAT promoter in mouse liver. (F) ChIP (top) for association of WT USF-1 and K237 USF-1 mutant to the FAS promoter in 293F cells. The promoter activity of the −444 FAS-Luc (bottom) in cells transfected with WT USF-1 or K237 USF-1 mutants was measured.