M1-Polarized Macrophages Promote Self-Renewing Phenotype of Hepatic Progenitor Cells with Jagged1-Notch Signalling Involved: Relevance in Primary Sclerosing Cholangitis

Figure 3

Classical macrophage activation is associated with elevated Notch ligand Jagged1 expression in macrophages and activation of Notch signalling in cocultured HPCs. (a) Quantitative analysis of Notch ligands (Jag1, Jag2, Dll1, and Dll4) expression in M0, M1, and M2 macrophages. (b) Jagged1 levels on M1 macrophages are confirmed by flow cytometry and Western blots. Flow cytometry results are represented by scatter plot, and calculated flow cytometry data are represented; vs M0 macrophages and vs M2 macrophages. (c) Histograms from flow cytometry analysis represent Jagged1 expressions in M1 macrophages from EGFP transgenic C57BL/6 mice cultured with or without HPC for 24 hours. (d) Graphs show relative mRNA expression levels of HES1 in HPC alone or in HPC cocultured with M0, M1, or M2 macrophages; levels of HES1 staining (Alexa Flour 568) in HPC were determined by static cytometry. Graphs show calculated relative expression of HES1 in HPC cocultured with M0, M1, or M2 macrophages (percentage of HPC culture alone). vs HPC alone, vs M0 macrophages cocultured with HPC, and vs M2 macrophages cocultured with HPC. (c) Representative Western blots showing HES1 protein levels in HPC after coculturing with M0, M1, or M2 macrophages.

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