Hematoxylin solution counterstaining

Tissue samples were fixed in 10% neutral buffered formalin or 4% paraformaldehyde (PFA) and embedded in paraffin. In some cases, frozen sections were made. Immunohistochemistry was performed as previously described (Zhu et al., 2010 (link)). Two hours before euthanasia, 50 mg/kg of BrdU (Sigma) was administered intraperitoneally. Primary antibodies used: Arid1a (Cell signaling, #12354s), Ki-67 (Abcam, ab15580), Cyp2e1 (Abcam, ab28146), and BrdU (BD 555627). Fibrosis detection was performed with Sirius red staining kit from Thermo Fisher Scientific (#NC0249910) or trichrome (UTSW Pathology Core). Apoptosis detection was performed with a Tunel kit from Roche (#11684817910). Detection was performed with the Elite ABC Kit and DAB Substrate (Vector Laboratories), followed by Hematoxylin Solution counterstaining (Sigma).

Tissue samples were fixed in 4% paraformaldehyde (PFA) and embedded in paraffin. In some cases, frozen sections were made. Immunohistochemistry was performed as previously described (Zhu et al., 2010 (link)). Primary antibodies used: Anti-Ki67, Anti-Cyp2e1, Anti-GS, Anti-Pten, Anti-Ctnnb1, Anti-Phospho-Histone H2A.X. For immunohistochemistry, detection was performed with the Elite ABC Kit and DAB Substrate (Vector Laboratories), followed by Hematoxylin Solution counterstaining (Sigma). For immunofluorescence, second antibodies Alexa Fluor 594 goat anti-mouse IgG1 or Alexa Fluor 488 goat anti-rabbit were used. Confocal images were taken by Zeiss LSM 780 Upright confocal/multiphoton microscope. H&E slides were interpreted by a clinical pathologist with expertise in human liver cancer diagnosis.