Detection of vaso-occlusion in Townes sickle mice following manipulation of PMo numbers. (A) Experimental design for manipulation of PMo numbers in Townes sickle mice with Clo-Lipo and MDP. Sickle mice (8-12 weeks) were injected IV with MDP daily from day 0 to day 3 or with Clo-Lipo at day 0 or with PBS daily from day 0 to day 3. Mice were sacrificed at day 5 for histological analysis. (B) Representative histograms showing the frequencies of PMos (CD45⁺CD11b⁺Ly-6G⁻CD115⁺Ly-6C⁻) and CMos (CD45⁺CD11b⁺Ly-6G⁻CD115⁺Ly-6C⁺) in 3 groups of sickle mice as defined in panel A. (C) The absolute number of circulating PMos, CMos, and neutrophils in 3 groups of mice (n = 6-7) as defined in panel A. (D) Representative H&E-stained liver sections in 3 groups of mice (scale bar, 200 µm). Black arrows indicate RBC stasis within blood vessels. Blue arrows indicate infarct. (E) Frequencies of blocked blood vessels (stasis) in total blood vessels in liver sections in 3 groups of mice (n = 6 in each group). (F) Frequencies of area of necrosis (infarct) in liver sections in 3 groups of mice (n = 6 in each group). (G) Representative immunofluorescence staining of liver sections from 3 groups of mice showing CD31/CD144 (endothelial markers, green) and Ter-119 (RBC marker, red). Scale bar, 50 µm. (H) Enumeration of Ter-119⁺ RBCs per image (0.18 mm² area) in liver sections in 3 groups of mice (n = 6-7 in each group) as quantified using ImageJ software. Data represent mean ± SEM. Means in panel C were compared using 2-way ANOVA, and means in panel F were compared using a 2-tailed Student t test. *P < .05; **P < .01; ***P < .001.