2.5. Histology, Picro Sirius red staining (PSR), and immunohistochemistry (IHC)

Paraffin-embedded liver tissue sections (5 μm thick) were deparaffinized in xylene and rehydrated and processed for hematoxylin and eosin and PSR (1.2% w/v; Direct red-80 Sigma-Aldrich) staining. Images were taken using an inverted light microscope (Zeiss Axiovert CFL 40 Trinocular Inverted Fluorescence Microscope). For IHC, tissue sections were baked at 80 °C for 15 min and rehydrated. Antigen retrieval was done by heating in the microwave with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH-6.0). Two different kits used for immunohistochemical staining; VECTASTAIN ABC KIT (Biotinylated Horseradish Peroxidase Anti-rabbit IgG) and ImmPRESS Duet Double staining Polymer kit (Horseradish Peroxidase Anti-Mouse IgG/Alkaline Phosphatase Anti-Rabbit IgG).