Figure 6.

Hepatic SHP stability is increased by CA feeding or FGF19 treatment in vivo and is abnormally elevated in metabolic disease model mice. (A) Mice were treated with vehicle PBS (−) or FGF19 (+) via the tail vein and, 6 h later, livers were collected for qRT–PCR. (B) Mice were injected with Ad-Flag-SHP and, 5 d later, fed normal control chow or 0.5% CA-supplemented chow (CA) for 14 h, or treated with FGF19 for 6 h. Nuclear and cytoplasmic extracts were prepared and Flag-SHP and lamin (as a nuclear protein control), tubulin (as a cytoplasmic protein control), and GFP (as a monitor of infection efficiency) were detected by Western analyses. Consistent results were observed from three sets of mice. (C) Mice were fed control chow or CA chow or were injected with FGF19 and livers were collected for Western analysis. (D) Livers from normal wild-type CV57 mice (N) or ob/ob mice were collected for qRT–PCR. (E) Wild-type normal (N) or ob/ob mice were injected with Ad-Flag-SHP and, 5 d later, cytoplasmic and nuclear extracts were prepared for Western blotting. Consistent results were observed from two sets of mice. (F) Livers from normal mice (N) or mice fed a high-fat and high-calorie western-style diet (WD) for 16 wk were collected for qRT–PCR. (G) Diet-induced obese mouse model: Livers from mice fed normal chow (N) or fed a chronic western diet (WD) for 16 wk were collected. Endogenous Shp levels were detected by Western analyses using SHP antibody. Results from two sets of mice are shown. (B,E,G) Band intensities were determined using ImageJ and the values for control or normal samples were set to 1. (A,D,F) Statistical significance was determined by the Student's t-test (SEM, n = 3). (**) P < 0.01; (NS) statistically not significant.