Figure 2. MoMF-derived JAG1 signaling is required for the generation of SOX9⁺ hepatocytes in the early stages of liver repair after injury.

(A) Representative immunofluorescent staining of SOX9 and β-catenin of liver tissues from ConA-treated mice (48 hours after treatment, n = 4). Red arrows indicate SOX9⁺ hepatocytes in the ConA-injured liver; yellow arrows indicate normal hepatocytes. β-Catenin shows membrane of hepatocytes. The average sizes of hepatocytes were quantified based on β-catenin staining. (B) Mice were treated with ConA for 24 hours, followed by injecting clodronate liposomes or control liposomes. Liver samples were collected 48 hours after ConA treatment. SOX9 and IBA1 double staining were performed on these samples (n = 5). (C) Mice were treated with ConA for 48 hours, followed by staining of liver tissues with JAG1 and SOX9 antibodies (n = 5). (D) Heterozygous CCR2-RFP mice (CCR2⁺ MoMFs are labeled with RFP) were treated with ConA. Liver MNCs were isolated and subjected to flow cytometry analyses of JAG1 and RFP (n = 5). (E) WT and hepatocyte-specific Notch1- and Notch2-knockout mice were treated with ConA for 48 hours. SOX9 protein in the liver tissues was stained, and the number of SOX9⁺ hepatocytes was quantified (n = 6). Representative images are shown in A, B, C, and E. Values in A, B, D, and E are represented as means ± SD. Statistical significance was assessed using 2-tailed Student’s t test for comparing 2 groups (B) and 1-way ANOVA followed by Tukey’s post hoc test for multiple groups (A and E). ***P < 0.001. Dashed lines indicate the borderlines of necrotic areas.