Analysis of liver tissue macrophages. 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. (A) Perfused liver cryosections were fixed, permeabilized, and incubated with anti-F4/80 (rat) and anti-CD31 (goat) antibodies followed by Alexa Fluor 562–conjugated anti-rat 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. (B) Perfused liver cryosections were fixed, permeabilized, and incubated with Clec4F (goat) and anti-CD31 (rat) antibodies followed by Alexa Fluor 562–conjugated anti-goat and Alexa Fluor 488–conjugated anti-rat antibodies. DAPI was used to stain the nucleus. Scale bar, 50 μm. (C-D) Relative intensity of F4/80 and Clec4F stain presented. All data 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. MFI, mean fluorescence density; ns, not significant.