Glycogen periodic acid schiff stain kit

TM was purchased from Cell Signaling (Danvers, MA, USA). Matrigel and collagen type I were purchased from Corning (Bedford, MA, USA). Human TGF‐beta1 Quantikine ELISA Kit was purchased from R&D Systems (Minneapolis, MN, USA). DAPI and pyrvinium were purchased from Sigma (St. Louis, MO, USA). Phalloidin was purchased from Yuheng Biotechnology (Suzhou, China). Glycogen Periodic Acid Schiff stain kit was purchased from Solarbio Life Sciences. Puromycin was purchased from BioFroxx (Guangzhou, China). X‐tremeGENE transfection reagent was purchased from Roche (Mannheim, Germany). Antibodies used were as follows: human CD44–FITC and CD24–phycoerythrin and their respective isotype controls were obtained from BD Biosciences (Franklin Lakes, NJ, USA). β‐Catenin was purchased from Santa Cruz (Dallas, TX, USA). VE‐cadherin, integrin β1, BiP, CHOP, Smad2/3, Phospho‐Smad2, Phospho‐β‐Catenin (Ser33/37/Thr41) and Phospho‐β‐Catenin (Ser675) were purchased from Cell Signaling. Bmi‐1 was purchased from Epitomics (Burlingame, CA, USA). Short hairpin RNA (shRNA) encoding β‐Catenin and scramble control shRNA were purchased from TsingKe Biological Technology (Chengdu, China).

Sorafenib (S7397, Selleck) and Ac‐PHSCN‐NH2 (ATN‐161) (S8454, Selleck) were solubilized in DMSO or sterile water with a concentration of 100 and 40 mM, respectively, stored at −20°C for further use. CoCl₂ (232696, Sigma) was weighed and solubilized directly in culture media for use. RPMI‐1640 medium, antibiotics (penicillin and streptomycin) and fetal bovine serum utilized in this study were obtained from Gibco (Grand). Hypoxyprobe™ Plus Kit was acquired from (HP2‐100Kit; Hypoxyprobe Inc). Glycogen Periodic Acid Schiff Stain Kit was purchased from Solarbio (G1281). We obtained all other analytical grade reagents from Fisher Scientific and Sigma‐Aldrich and used them without further purification.

For H&E staining, the 5‐μm sections were dewaxed and hydrated, then stained with haematoxylin and eosin (H&E) reagents. It was applied to evaluate the histological changes in liver tissues. For PAS staining and Masson's staining, after the 5‐μm sections were dewaxed, Glycogen Periodic Acid‐Schiff Stain Kit (Solarbio, G1281) and Masson's Trichrome Stain Kit (Solarbio, G1340) were used to stain the liver tissues according to the instructions to evaluate the levels of glycogen and collagen in the liver tissues. Then, the images were observed and captured using a Nikon ECLIPSE 80i microscope (Nikon, Japan).