The right medial, the left lateral, and the caudate lobe of the liver were collected for histological examination. Samples were fixed in 10% Neutral Buffered Formalin, processed to paraffin, imbedded and cut in 2–4 µm sections. In addition, a sample from the left medial lobe was snap frozen for cryo-sectioning. Steatosis was evaluated in all three lobes with Mayer’s Haematoxylin and Eosin (H&E) on paraffin section (10 animals/gr) and confirmed by Oil Red O-stain on frozen cryo-sections of the left medial lobe (2–3 animals/group). Inflammation and fibrosis were evaluated based on the morphology on the H&E stain, collagen deposition on a Picro Sirius stain and macrophage infiltration based on a CD68 immunohistochemistry (IHC) stain. For the CD68 IHC stain, antigens were first retrieved by TEG buffer pH 9.0 and subsequently blocked with 0.5% hydrogen peroxide followed by avidin and biotin (Invitrogen, CA, USA). The slides were then incubated with primary antibody (3 µg/mL, MCA341R, AbD serotec, CA, USA) for 60 min, biotinylated secondary antibody for 60 min (1 µg/mL, 715-065-151, Jackson ImmunoSearch, PA, USA) and ABC reagent (Vector Laboratories, CA, USA). Finally, macrophages were visualised with a DAB reagent (Dako, Glostrup, Denmark). All sections were scanned with a Hamamutsu slide scanner and later evaluated using NanoZoomer Digital Pathology Image software (Hamamatsu, Hamamatsu City, Japan). The collagen and macrophage areas were quantified by Visiomorph software (Visiopharm, Hoersholm, Denmark).