Figure 2.

Tb administration down-regulates hepatic CPT-1A expression in mice. Mice were pair-fed (isocaloric maltose dextrin [PF]) with either a control diet or EF (5% wt/vol) along with Tb (2 g/kg, EF + Tb) administered orally. (A) CPT-1A mRNA levels were analyzed by real-time quantitative PCR and normalized to β-actin mRNA. (B) Immunohistochemical staining with anti–CPT-1A antibody (original magnification, 20×). Representative images are shown. Bar graph shows the quantification of CPT-1A protein using the MetaMorph Microscopy Automation and Image Analysis Software (Molecular Devices, LLC, San Jose, CA) by calculating the percentage of positive (based on the average intensity of the field-of-view) microscope fields. Detection of CPT-1A protein levels by Western blot analysis in mice liver tissues. Blots were stripped and re-probed with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody as a loading control. The density ratio for each band compared with its GAPDH is shown as a bar graph. Data are expressed as means ± SEM, n = 5 animals/per group. P value <.05 (a) when compared with PF, (b) when compared with EF. (C) ChIP analysis was performed on mice liver tissue. Chromatin fragments were immunoprecipitated with anti-H3K9Ac antibody and ChIP-qPCR was performed using primer pairs specific for the TRE region on the CPT-1A promoter. Nonimmunoprecipitated chromatin was used as input. Data were analyzed by analysis of variance and represented as means ± SD, n = 4–6 mice per group (n = 3 for Western blot). P value < .05 (a) when compared with PF, (b) when compared with EF.