Fig. 5

Transplantation of CD24⁺LCN2⁺ LPCs exacerbated chronic fibrosis and inflammation by infiltrating macrophages. a Schematic overview of the experimental design. Eight-week-old wild-type C57BL/6 mice were injected with 2 mL/kg CCl₄ i.p. twice a week for a continuous period of 6 weeks. 1–2 × 10⁶ CD24⁺LCN2⁺ LPCs or HepLPCs were transplanted 4 h after the first CCl₄ injection. b Representative images of H&E, Sirius Red, and immunofluorescence staining for CK19, α-SMA, and F4/80 in liver slides. Scale bar: 600 μm for H&E staining and Sirius Red staining; 200 μm for immunofluorescence staining. c Quantification of Sirius Red positive area as a proportion of the total liver area. Data were measured by Image J software. d Serum analysis of ALT and AST. e Circle plots showing the number of interactions between CD24⁺LCN2⁺ LPCs and other cell groups in NASH-NPCs of Fig. 4. f Violin plots showing expression of Il1b, Tnf, and Cd86 in NPCs of Fig. 4. g Schematic model of the transwell assay and quantification of migrated cells in basal medium, CD24⁺LCN2⁺ LPC conditioned medium or HepLPC conditioned medium. h Quantification of the cell number at the bottom of transwells. i RT-q-PCR analyses M1 macrophage expression. M0 macrophages were cultured in basal medium (BM) or conditioned media (CM) of CD24⁺LCN2⁺ LPCs or HepLPCs, as indicated. j RT-q-PCR analyses of M1 macrophage expression. M1 macrophages were cultured in basal medium (BM) or conditioned media (CM) of CD24⁺LCN2⁺ LPCs or HepLPCs, as indicated. k, l Quantification of the cell number at the bottom of transwells k and RT-q-PCR analyses of M1 macrophage expression l M0 macrophages were cultured in basal medium (BM) or CD24⁺LCN2⁺ LPC-conditioned mediums with or without Lcn2 expression. Resident LPCs-derived CD24⁺LCN2⁺ LPCs, reLPCs; Hepatocytes-derived LPCs, HepLPCs. For panels c, d, h, i, j, k, and l, data are expressed as means ± SD of three independent experiments. p values were analyzed by one-way ANOVA. *p < 0.05; **p < 0.005; ***p < 0.001; ns: no significant difference