Reduced triglyceride accumulation in PASK−/− livers on HFD. (A) Representative oil red O staining of liver cryosections from 24-week-old WT and PASK−/− male mice on NCD and HFD after an 8-h fast. (B) Liver triglyceride (triacyl glycerol, TAG) content in 24-week-old WT and PASK−/− mice on NCD and HFD harvested after an 8-h fast (n = 3–5 male mice). (C) Immunoblot analysis of phospho-AMPK (Thr-172), phospho-S6K (Thr-389), and tubulin (as loading control) levels in liver extracts from WT and PASK−/− mice on NCD and HFD. Results from two mice per group are shown. Total AMPK and S6K protein levels were also analyzed and found to be identical among the groups. (D) SCD1, fatty acid elongase (FAE), CD36, PPARγ, CYP3A11, and PXR mRNA levels in liver from WT and PASK−/− mice on HFD measured by quantitative RT-PCR. The data shown are the average of n = 6 per group with the WT value set as 1. RPL13A was used as the normalizer. All data presented are the mean ± SD.