(A) Schematic representation of experimental design: mice were treated with an agonistic anti-CD40 antibody for 30 hours (for macrophage isolation and blood removal) or 48 hours (for liver collection). (B) Representative histology image for H&E staining of a thrombus with an ischemic liver infarct after treatment of Spta^wt/wt mouse with anti-CD40 antibody. Scale bar: 200 μm. (C) Plasma alanine transaminase (ALT) concentrations in LysM-Cre Cd40^fl/fl (Cre^+/–, knockout) or Cd40^fl/fl (Cre^–/–, control) mice treated with saline or anti-CD40 antibody (n = 3–6). (D) Plasma ALT concentrations in Spta^wt/wt (WT, blue) and Spta^sph/sph (Sph, red) mice treated with saline or anti-CD40 antibody (n = 6–8). (E) Photographs of representative liver lobes from Spta^sph/sph and Spta^wt/wt mice treated with anti-CD40 antibody. (F) Hierarchical clustering analysis of plasma cytokines from Spta^sph/sph and WT littermates treated with anti-CD40 antibody (yellow = low concentration, red = high concentration) or saline. (G) Bulk RNA-seq data of F4/80-enriched nonparenchymal liver cell suspensions from 3 Spta^sph/sph and Spta^wt/wt mice treated with anti-CD40 antibody or saline. Hierarchical clustering analysis of the 2000 top significantly DEGs (rows) between the anti-CD40 and saline-treated Spta^wt/wt (column). In total, 5803 DEGs were identified based on an absolute log₂(ratio) > 0.5 and P value < 0.01 threshold. (H) GSEA of DEGs of KCs in Spta^wt/wt mice treated with anti-CD40 antibody versus saline. Enrichment plots of the top 4 positively enriched hallmark gene sets are shown. Plots display running enrichment score and position of gene set members on the rank-ordered list. (I) Normalized count data for key inflammatory genes expressed in KCs from anti-CD40– or saline-treated (cont) Spta^wt/wt (blue) and Spta^sph/sph (red) mice. Each data point represents a single mouse. ***P < 0.001; **P < 0.01 by ANOVA with Tukey’s post hoc test for C and D; counts were normalized using DEseq2 for I.