Fig. 4.

FXR and FoxO3a did not interact in vitro or in vivo. HEK 293A cells were transfected with plasmids containing flag-FXR, HA-FoxO3a, HA or flag. Total cell lysates were isolated, and flag was pulled down by immunoprecipitation and subjected to immunoblot analysis for flag and HA. Input total lysates from transfected HEK 293A were used as positive controls for FoxO3a and FXR (A). Cytosol and nuclear fractions were obtained from transfected HEK 293A cells, and immunoblot analysis was performed for flag and FoxO3 (B). Age matched WT and FXR KO mice were treated with 4.5 g/kg ethanol or water by gavage for 16 h. FoxO3a in total liver lysates was pulled down by immunopreciptation and subjected to immunoblot analysis for FXR and FoxO3a (C).