Fig. 1

Characterization of immortalized hepatocyte-like cells (imHCs). imHCs were maintained in DMEM/F12 medium containing 10% fetal bovine serum. After reaching confluence, cells exhibited a typical hepatocyte morphology, including a polygonal shape, granulated cytoplasm, and large nucleus (a). Hepatocyte phenotypes were evaluated via immunofluorescence staining for the following major hepatocyte markers: albumin (ALB) (b), ɑ-fetoprotein (AFP) (c), hepatocyte nuclear factor-4-alpha (HNF-4ɑ) (d), low-density lipoprotein receptor (LDLR) (e), and Na⁺-taurocholate cotransporting polypeptide (NTCP) (f). Cell nuclei were visualized using Hoechst 33342 DNA dye. Fluorescence images were captured and analysed using an Operetta High-Content Imaging System (PerkinElmer) with a ×40 objective lens. Scale bar = 50 μm. The presence of hepatic marker in imHCs was quantified from 15 randomly selected image fields (total number of analysed cells > 2000) (g). Columns represent the percentage of cells containing ALB, AFP, HNF-4ɑ, LDLR or NTCP. Data are represented as mean ± standard deviations