Figure 5.

Expression profiling in liver and skeletal muscle of Stk25 knockout mice and wild-type littermates challenged with a high-fat diet for 20 weeks. A and D: Protein lysates of the liver (A) and gastrocnemius skeletal muscle (D) were analyzed by Western blot using antibodies specific for total ACC, phospho-ACC (Ser⁷⁹), total AMPKα, phospho-AMPKα (Thr¹⁷²), or STK25. Protein levels were analyzed by densitometry, and data are shown as the total and phospho-protein abundance as well as the ratio of phospho-protein to total protein, with values in wild-type mice set to 1. Representative Western blot is shown with actin (A) or glyceraldehyde-3-phosphate dehydrogenase (D) used as loading controls. B: Protein lysates of liver were enriched for ubiquitinated proteins followed by Western blot using antibodies specific for total ACC. Amount of ubiquitinated ACC was analyzed by densitometry, and the level in wild-type mice was set to 1. C and E: Relative mRNA expression assessed by quantitative real-time PCR in the liver (C) and gastrocnemius skeletal muscle (E). The expression level of each selected gene in wild-type mice was set to 1. The gene functions are indicated at the bottom. Data are mean ± SEM from 7–10 mice per genotype. *P < 0.05, **P < 0.01 for Stk25^−/− mice vs. wild-type littermates. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HFD, high-fat diet; KO, knockout; p, phospho; Ub-ACC, ubiquitinated acetyl-CoA carboxylase; WT, wild type.