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

Figure 4

Blocking Jagged1 in a coculture system attenuates hepatic progenitor cell self-renewal as well as HES1 expression in HPC. (a) HPCs were cocultured (24 h) with M1 macrophages from EGFP transgenic C57BL/6 mice with or without the existence of anti-Jagged1 mAb (10 μg/ml). Graphs show fluctuation of CD44⁺CD324⁺ and CD44⁺CD326⁺ cell fractions in HPC cultured alone, cultured with M1 macrophages or with anti-Jagged1 mAb in coculture system. (b) Representative Western blots showing protein levels of Jagged1 in M1 macrophages transfected with mock or siJag1. In the cases that macrophages were transfected with mock or siJag1 vectors before M1 polarization, CD44⁺CD324⁺ and CD44⁺CD326⁺ cell fractions of HPC are highlighted by calculated histogram. (c) Graphs and representative Western blots showing protein levels of HES1 and mRNA expression of HES1 in HPC when cocultured with M1-siJag1 or cocultured with M1 macrophages and 10 μg/ml anti-Jagged1 mAb. vs HPC alone, vs M1 macrophages cocultured with HPC + anti-Jagged1 antibody, and vs M1 macrophages cocultured with HPC + siJag1.

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