Periodic acid schiff stain kit

Reagents were purchased from Sigma-Aldrich (St. Louis, MO, United States), unless otherwise indicated. Antibodies against phospho-moesin (T558), moesin, and CD34 were purchased from Abcam (Cambridge, UK). β-actin antibody was obtained from Cell Signaling Technology (Beverly, MA, United States). Biotin-free horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Sigma (St. Louis, MO, United States). Tyramide-CY3 and FITC were obtained from Servicebio (Wuhan, China). DAPI was obtained from BestBio (Shanghai, China). RIPA Lysis Buffer, PMSF, phosphatase inhibitor cocktail, and bovine serum albumin (BSA) were all obtained from GBCBIO (Guangzhou, China). Hematoxylin and Eosin (HE) Stain Kit, Periodic Acid Schiff (PAS) Stain Kit, and Masson’s trichrome Stain Kit were purchased from Solarbio (Beijing, China).

Oil red O staining: Fresh liver tissues were fixed with 4% paraformaldehyde and then prepared as 10 μm thick frozen sections. Air-dried frozen sections were soaked in distilled water for 10 min and then briefly rinsed with 60% isopropanol. After being stained with freshly prepared oil red O working solution for 10 min, sections were rinsed with 60% isopropanol. Following by distilled water washing, sections were stained with Mayer's hematoxylin for 10 s. Afterward, the stained sections were thoroughly washed in running water for 5 min. Finally, before image acquisition, the sections were rinsed with distilled water and mounted with glycerin.
Periodic acid Schiff (PAS) staining: The wax embedded liver tissues were prepared as 3 μm thick paraffin sections for glycogen detection. Liver glycogen was detected using a Periodic Acid Schiff Stain Kit (G1281, Solarbio, Beijing) and the assay was executed according to the manufacturer's instructions.

A Periodic Acid Schiff Stain Kit (Solarbio, Beijing, China) was used to show the PAS-positive laminated layer characteristic of E. multilocularis metacestodes. The section (5 μm) was dewaxed in xylene and rehydrated in 100%, 95%, 80%, and 75% alcohol baths. Staining was then carried out according to the kit instructions.

Immunofluorescence staining of hepatocyte was performed as that described for iPSC characterization. The primary antibodies used were anti-SOX17 (R&D SYSTERM #AF 1924, Minneapolis, MN, United States), anti-FOXA2 (Merck Millipore #07–633), anti-FVIII N-terminus antibody (Santa Cruz Biotechnology #sc27649, Dallas, TX, United States), and anti-AFP (Sigma-Aldrich #A8452), anti-ALB (R&D SYSTERM #MAB1455).
For periodic acid schiff stain, the hepatocytes on Day 25 were stained with the Periodic Acid Schiff Stain Kit (Solarbio #G1280, Beijing, China) according to the manufacturer’s instructions. The periodic acid schiff stain was detected by microscopy.
For indocyanine green (ICG) uptake assay, hepatocytes on Day 25 were treated with 1 mg/mL ICG (Sigma-Aldrich #1340009) for 30 min, then washed with DPBS thoroughly and cultured in fresh Hepatocyte Culture Medium. The cells were detected by microscopy. After 12 h, the cells were observed using microscopy.