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

Figure 6. HNF-4α physically interacts with ChREBP in vivo.

Figure 6

(A) HEK-293 cells were co-transfected with expression vectors for HNF-4α and FLAG-tagged ChREBP. Nuclear protein extracts were prepared, 42 h following transfection, and subjected to co-immunoprecipitation with anti-HNF-4α antibody. After incubation at 4 °C overnight and washing three times with PBS containing 0.1% Nonidet P40, the samples were analysed on a 10% gel by SDS/PAGE and Western blotted with anti-FLAG antibody. The top and middle panels are Western blots of 5% of the nuclear extract before immunoprecipitation, and the bottom panel is a Western blot of the immunoprecipitated proteins. (B) Nuclear protein extracts from the liver of refed rats were prepared, subjected to immunoprecipitation with anti-HNF-4α antibody or preimmune IgG (PI), and Western blotted with anti-ChREBP antibody (top panel). The blot was then stripped and immunoblotted with a separate anti-HNF-4α antibody (bottom panel). (C) The inverse of the experiment displayed in (B) was performed, with nuclear protein extracts subjected to immunoprecipitation with anti-ChREBP antibody, and immunoblotted with anti-HNF-4α antibody (top panel). The blot was stripped and immunoblotted with anti-ChREBP antibody (bottom panel).