Journal: Stem cell research & therapy

Article Title: Hepatic progenitor cells reprogrammed from mouse fibroblasts repopulate hepatocytes in Wilson's disease mice.

doi: 10.1186/s13287-025-04253-1

Figure Lengend Snippet: Fig. 1 Coexpression of Foxa3 and Hnf4α reprogrammed mouse fibroblasts into iHPCs. (A) Constructs of the control-lentiviral vector and the overexpres sion-lentiviral vector carrying EF1α-promoter-driven Foxa3 and CMV-promoter-driven Hnf4α. (B) Representative phase-contrast morphology of control fibroblasts and iHPCs on Days 0, 2, 7, and 14 after lentivirus infection. Scale bars, 100 μm. (C) Immunofluorescence staining Flag and DAPI in control fibroblasts and iHPCs. Scale bars, 50 μm. (D) Immunofluorescence staining and flow cytometry were used to analyse the percentages of Flag-positive control fibroblasts and iHPCs. (E) Real-time PCR was used to determine the relative expression of Foxa3 and Hnf4α in control fibroblasts and iHPCs. (F) Western blot analysis showing the relative protein levels of Foxa3, Hnf4α, Alb, Cdh1, and β-Actin in control fibroblasts and iHPCs. β-Actin served as a load ing control. (G) Giemsa staining (scale bars, 50 μm), PAS staining (scale bars, 25 μm), and phase-contrast morphology of control fibroblasts and iHPCs at serial passages (scale bars, 100 μm)

Article Snippet: After being blocked with TBST buffer containing 5% nonfat dry milk, the membranes were incubated with one of the following primary antibodies: Hnf4α (Cell Signaling Technology, Danvers, MA, 1:1000), albumin (Alb, R&D Systems, Minneapolis, MN, 1:1000), E-cadherin (Cdh1, Abcam, Cambridge, UK, 1:1000), Foxa3 (Abcam, 1:1000), or β-Actin (Abcam, 1:1000).

Techniques: Construct, Control, Plasmid Preparation, Infection, Immunofluorescence, Staining, Flow Cytometry, Positive Control, Real-time Polymerase Chain Reaction, Expressing, Western Blot