Skip to main content
. 2024 Jun 13;11(30):2403516. doi: 10.1002/advs.202403516

Figure 1.

Figure 1

Distinct circulating immune cells are mobilized to the liver and modulate hepatocyte responses to APAP and FFAs. A) Archival formalin‐fixed paraffin‐embedded mouse liver samples were retrieved and subjected to multiplex immunohistochemistry. B) Digital image segmentation and analysis were performed. Heatmaps represent immune cell location in the respective mouse models. C) Imaging cytometry allowed for immune and pathology‐associated cell numbering, and individual cell staining intensity measurements. Individual data points are displayed in Figures S1D,  S2D (Supporting Information. D–I) Conventional 2D cell culture was performed using primary mouse hepatocytes and CICs, exposed to either APAP or FFAs. D) Immunocytochemistry was performed to visualize cell death. Quantification is shown in (E). F) Cells were cultured in the presence of FFAs and BODIPY staining was applied to visualize lipid deposition. G) BODIPY quantitation. H,I) Relative gene expression was analyzed by qRT‐PCR. Sample sizes: A–C) n(Veh) = 4, n(APAP) = 4, n(control diet) = 4, n(CDAHFD + Veh) = 7, n(CDAHFD + Lanif) = 7. D–I) n = 4 per group. Abbreviations: APAP: acetaminophen; CDAHFD: choline‐deficient, amino acid‐defined high‐fat diet; Lanif: lanifibranor; Veh: vehicle; CK7: cytokeratin 7; CD3: a cluster of differentiation 3; CD45R: cluster of differentiation 45 receptor; IBA1: Ionized calcium‐binding adaptor molecule 1; Ccl: Chemokine (C‐C motif) ligand; CLEC4F: C‐Type Lectin Domain Family 4 Member F; MPO: myeloperoxidase; PCNA: proliferating cell nuclear antigen; CD11b: cluster of differentiation 11b; PDGFRβ: platelet‐derived growth factor receptor beta. C) One‐way ANOVA followed by Tukey's multiple comparison was performed. &p < 0.05; &&p < 0.01; &&&p < 0.005 as compared to vehicle; **p < 0.01; ***p < 0.005 as compared to control diet; §p < 0.05; §§p < 0.01; §§§p < 0.005 as compared to CDAHFD + vehicle. E,G,H,I) One‐way ANOVA followed by Tukey's multiple comparison test was performed. H, I) *p < 0.05 as compared to a vehicle; $p < 0.05 as compared to Veh + CIC.