PRN473 reduces Salmonella-induced liver thrombosis in mice. (A-B) WT mice were fed control or PRN473 formulated diet for 7 days, then had spleens harvested for Btk occupancy analysis (A) or citrated whole blood taken for platelet activation analysis by flow cytometry (B), indicated by activated integrin αIIbβ3 (JON/A) and P-selectin surface expression, after stimulation with snake venom toxin rhodocytin (100 and 300 nM), CRP (3 and 10 μg/mL), or PAR4 receptor activating peptide (500 nM). Mean ± SEM (n = 3). Statistical analysis by 2-way ANOVA with Sídák correction for multiple comparisons. (C) WT mice fed control or PRN473 formulated diet were infected with 5 × 10⁵ CFU S typhimurium on day 7, with thrombi in portal vein assessed at day 14 (7 days after infection). Representative immunohistochemistry staining of frozen liver sections (scale bar, 200 μm) (i) and quantification of thrombus area per unit vessel area (ii), quantification of number of thrombi (iii), quantification of number of podoplanin-expressing vessels (iv), number of thrombi in podoplanin-positive levels (v) (n = 6). (D) Peripheral blood counts of S typhimurium–infected mice at baseline and 7 days after infection. Mean ± SEM (n = 4). (A-B) ∗P < .05; ∗∗P < .01; ∗∗∗∗P < .0001. CFU, colony forming units; WT, wild type.