2.3 Histological and immunofluorescence staining
After fixation in formalin for one week, the livers were embedded in
paraffin and cut into 4.5 μm thick sections for subsequent staining.
After deparaffinization in xylene and dehydration in ethanol, the
sections were stained using a hematoxylin and eosin (H & E) staining
kit and a Masson’s staining kit. For immunofluorescence, the sections
were blocked with 0.2% Triton X-100-2.5% BSA-1 × phosphate-buffered
saline (PBS)-10% goat serum after subjecting to antigen retrieval
solution. The primary antibodies against FN (dilution 1:200), α-SMA
(dilution 1:400), COLLAGEN (dilution 1:400), F4/80 (dilution 1:300),
CXCL16 (dilution 1:360), CD68 (dilution 1:400), MPO (dilution 1:400),
citrullinated histone H3 (CitH3) (dilution 1:400), CD11b (dilution
1:400), α-SMA (dilution 1:400) were respectively incubated overnight at
4°C. After washing, the sections were incubated with goat anti-mouse IgG
(H+L) highly cross-adsorbed 488 secondary antibody or anti-rabbit IgG
(H+L) 594 secondary antibody (Thermo Fisher Scientific, Waltham, USA)
and stained with DAPI. After staining of cell nuclear, the sections were
sealed with resin and further observed using Aperio Versa (Leica,
Wetzlar, Germany).