Ab190958

Mouse livers were collected at the specified time points, fixed with 10% neutral buffered formalin and embedded in paraffin for processing into 4 micron sections. Sections were rehydrated and stained with Haematoxylin and Eosin (H&E) (Thermo Scientific) or Sirius red and Fast green (Sigma) before pathology evaluation. Alternatively, sections were treated with heat mediated antigen retrieval with pH6 sodium citrate or pH9 Tris-EDTA buffer before staining with primary antibodies. Primary antibodies used in this study include anti-HSA (ab2406, ab19180 Abcam), anti-CD3 (ab699, Abcam), anti-CD20 (555677, BD), anti-CD68 (ab955, Abcam), anti-PD1 (ab137132, Abcam), anti-β-catenin (610154, BD), anti-glutamine synthetase (MAB302, Millipore), anti-LFABP (ab190958, Abcam), anti-IL6 (ab6672, Abcam) and anti-IL10 (ab34843, Abcam).
Immunohistochemistry staining was performed using the Superpicture 3^rd Gen IHC Detection Kit (879673, Life Technologies) while immunofluorescence staining was done using anti-mouse IgG, anti-rabbit IgG or anti-Goat IgG secondary antibodies conjugated to Alexafluors 488 or 647 (Life Technologies). Images were captured using an Olympus upright confocal microscope or a Zeiss Axioscan Z1 slide scanner.

Protein extracts were prepared from the thawed liver, heart, and kidney tissue samples. Equal amounts of proteins were separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (TGX™ FastCast™ Acrylamide Kit, 12% NO. 1610175). The tissues were then transferred onto polyvinylidene difluoride (PVDF) membranes (Trans-Blot® Turbo™ Mini PVDF Transfer Packs NO. 1704156; Bio-Rad, USA) and were subsequently blocked with nonfat dry milk (Blotting-Grade Blocker NO. 1706404 for western blot applications). The membranes were incubated with primary antibodies against PPAR-α (ab24509; Abcam, USA), PPAR-γ (ab209350; Abcam, USA), CPT1A (ab83862; Abcam, USA), liver FABP antibody-N-terminal (ab190958; Abcam, USA), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (ab181602; Abcam, USA). After washing with 0.1% Tween 20 in Tris-buffered saline (TBS), the membranes were incubated with the horseradish peroxidase-conjugated secondary antibody (ab6721; Abcam, USA). The images were detected by the ChemiDoc MP System imager. The bands were quantified and analyzed by JLab software.