Western blot analysis of signaling molecules and infiltration of GR1-positive leukocyte to the liver tissues. Mice were treated with control siRNA and AT siRNA, followed by infusion of saline, AT-WT, AT-4Mut, or AT-R425del at 24 and 48 hours of siRNA treatment. Mice were euthanized after 72 hours of siRNA treatment and perfused with PBS. (A) The perfused left lobe of the liver was harvested in the tissue lysis buffer and immunoblotted for VCAM1, myeloperoxidase (MPO), and RAGE. β-actin was used as a loading control. (B-D) Densitometric analysis of VCAM1, MPO, and RAGE expression in liver tissue samples. (E) The perfused left lobe of the liver was harvested in the tissue lysis buffer and immunoblotted for VCAM1, MPO, and RAGE expression in liver tissue samples. β-actin was used as a loading control. (F-H) Densitometric analysis of VCAM1, MPO, and RAGE in liver tissue samples. (I-J) Mice were treated with control siRNA or AT siRNA followed by infusion of saline, AT-WT, AT-4Mut, or AT-R425del at 24 and 48 hours of siRNA treatment. (I) Perfused liver cryosections were fixed, permeabilized, and incubated with anti-GR1 (rat) and anti-CD31 (goat) antibodies followed by Alexa Fluor 555–conjugated anti-rabbit and Alexa Fluor 488–conjugated anti-goat antibodies. DAPI was used to stain the nucleus. Inset boxes from each group are magnified. Scale bar, 50 μm. (J) Number of GR1-positive leukocytes per field is presented. All data are presented as mean ± SEM. n ≥ 4. Statistical analysis was performed using Graph Pad Prism 10. P values are determined by 1-way ANOVA, followed by Bonferroni multiple comparison test. ∗P < .05; ∗∗P < .01; ∗∗∗P ≤ .001. ns, not significant.